OUTLINE

I.

Introduction

II.

Overview of the Animal Cell

III.

Considerations in Cell Development

IV.

Approaches to Biomedical Study

V.

TISSUE CULTURE

VI.

Stem Cell Research

VII.

References

I
INTRODUCTION
Preamble
It is a biological axiom that life is mediated by cells (Cooper, 2000). From life forms consisting of a single
cell, to the most fully developed form of complex multicellular organisation encountered in higher animals
which features separate and dependent levels of complex organisations of some 100 trillion cells into four
basic tissue types, which are further formed into distinct organs, and thence into organ systems (Guyton
and Hall, 2005), finally discharging their functions in pseudo-industrial organic complexes of
supersystems to sustain the existence of the organism that sits atop this biological edifice the cell governs
the complex biochemical interactions responsible for the phenomenon we understand to be life.

All cells share common fundamental properties that have been conserved throughout evolution. For
example, all cells employ DNA as their genetic material, are surrounded by plasma membranes, and use the
same basic mechanisms for energy metabolism. However, present-day cells have evolved a variety of
different lifestyles (Cooper, 2000). In unicellular organisms, all vital processes occur in a single cell, but
these processes in more complex organisms are distributed among specialised cells organised into
functional groupings along a gradient of increasing complexity, as already noted (Ganong, 2005).
Evolutionary theory posits that the evolution of multicellular organisms progressed by various cell groups
taking over particular functions and developing the specialized complexity to achieve this (Alberts et al,
2002, Cooper, 2000).

This organisational complexity has however been found to not be without its vulnerabilities. Chief of
which is the susceptibility of cells to damage, majorly from the fact of their own aggregated metabolism
(wear and tear) as well as the products of such metabolism. Cells have a basic ability to repair themselves
in the presence of limited damage, signalled by the control mechanism of the cell DNA and interaction
with its extracellular matrix. However, damage beyond a certain level impairs this communication, and
certain adaptations ensue. These adaptations are the cells physiological adjustment, a backup system as
it were, to ensure that the mission of the cell is continued to be prosecuted while avoiding greater damage.
Unfortunately, these physiological adaptations are in fact themselves pathological states, or result into one
when the cell attempts to operate under such alien conditions (Kumar et al, 2005).
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Remarkably, the cell is able to adjust even to these pathological states, up to an extent, after which its
function becomes completely aberrant. This aberrance may result in an ordinary organic impairment or it
may become neoplastic in nature. Or, more pernicious, it may affect the cell control structures (i.e. genetic
material and protein processing organelles) and this affects the development of subsequent cell lines. This
latter development is responsible for a number of life-threatening conditions with impairment of processes
vital to the organisms survival. Such an eventuality appears to have been envisaged during the
evolutionary process such that there inheres in the organism (in special storage) a reservoir of
undifferentiated primary type of cells which may be called upon to do duty to replace worn-out tissue, or
badly malformed ones, though in the ordinary course of things they do not do this (except in very
specialised tissues, or if they are made to do so). These special cells in reservoir are known as stem cells
(International Society for Stem Cell Research, 2016). Stem cells are so-called because they have the
remarkable potential to develop into many different cell types in the body during early life and growth. In
addition, in many tissues they serve as a sort of internal repair system, dividing essentially without limit to
replenish other cells as long as the person or animal is still alive. When a stem cell divides, each new cell
has the potential either to remain a stem cell or become another type of cell with a more specialized
function, such as a muscle cell, a red blood cell, or a brain cell (International Society for Stem Cell
Research, 2016; National Institutes of Health, 2015).

It must be pointed out that all cells of the early embryo are necessarily stem in nature (Yu and Thomson,
2006) otherwise no human could ever develop from a zygote. Three levels of stem are described in
human development, totipotency, pluripotency and multipotency. Cells up to the eight-cell stage of the
zygotes early division (cleavage) possess totipotence as each, isolated, can develop into a viable embryo
(Weiss et al, 2013). Pluripotence, on the other hand, is the ability of a cell to develop into any type of body
tissue. It used to be thought that pluripotence and totipotence were synonymous until recent insights
corrected that notion (Sadler, 2003). Pluripotence is now known to be the lesser totipotence (pluripotent
cells cannot develop to a viable embryo) and once differentiation commences in the human embryo (from
Day 5 post-fertilisation), the remainder undifferentiated cells lose totipotence to become only pluripotent.
These two properties do not remain long in the primordial cells, and once implantation occurs, the various
cells become disposed into layers and they slowly become specialised to region-, then tissue-specific cells
as they become dedicated to developing particular body tissues and organs (Yu and Thomson, 2006).
By the end of the embryonic period (8th week), when much of the bodys primordial developmental
template has been laid down, the resulting foetal cells retain only the property to divide and differentiate
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into different types of cells which is necessary to the proliferation of the tissues of specific organ systems.
This referred to as multipotency, and it is deemed the least type of stem. A very small population of these
are often buried deep within the developing tissue as cell reservoirs. Recent advances though have found a
use for these type of cells, and it is highly considered in many quarters to be the future of stem cell research
(International Society for Stem Cell Research, 2016). By the time the foetus comes to term and parturition
occurs, this is the stock of stem cells (by now solely multipotent in nature) to be found mainly in tissue and
organ depots in the organism, such as the bone marrow (the site of haemopoiesis) and adipose tissue. These
are the sites where specialised production of cells which have undergone terminal specialisation take place.
Their general occult nature makes them difficult to identify, isolate and grow in a laboratory setting.

This fact of the availability of stem cells, coupled with advances in biomedical research techniques, led
researchers to seek to actively influence, even to manipulate, the course of tissue repair by the use of stem
cells. And further, to bring about organ repair and renewal (and even replacement). This particular
endeavour, or rather conglomeration of research activities under the broad label of Stem Cell Research (due
to the one single strand that links all the various disparate efforts), has met with varying degrees of success,
and has weathered a great storm of challenges especially unrelenting criticism mainly on ethical and
religious grounds. Many countries of the world ban all types of stem cell research outright, while varying
degrees of restrictions are instituted on the scope of stem cell research that may be done in the countries
where it is legal to do stem cell research. This has affected the pace of discovery and development that
might otherwise have been had. A most potent polemic weapon has been to equate freedom to do stem cell
research with an open season of unrestrained assault on basic human dignity. This means translates to a
potential backlash with dire political consequences that policymakers in many free polities of the
contemporary world tread cautiously around this issue.

Statement of Purpose
This term paper will examine the concept of the stem cell, starting from a background of eukaryotic cells,
and attempt to provide a timeline of developments in this exciting and promising area of biomedical
science, while commenting on recent developments in the field. Biomedical research techniques will also
be examined, especially the Tissue Culture which has facilitated stem cell research in no small measure,
and is in fact the principal technique by which stem cell research is prosecuted (Yu and Thomson, 2006;
Zurlow et al, 1994).

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II
OVERVIEW OF THE ANIMAL CELL
All Cells are divided into two main classes, initially defined by whether they contain a nucleus, namely
prokaryotic cells (bacteria) and eukaryotic cells (Cooper, 2000). Prokaryotic cells (bacteria) lack a nuclear
envelope while eukaryotic cells have a nucleus in which the genetic material is separated from the
cytoplasm. Prokaryotic cells are generally smaller and simpler than eukaryotic cells. In addition to the
absence of a nucleus, their genomes are less complex and they do not contain cytoplasmic organelles or a
cytoskeleton. In spite of these differences, the same basic molecular mechanisms govern the lives of both
prokaryotes and eukaryotes (Alberts et al, 2002; Cooper, 2000).

A typical prokaryote cell

Three-dimensional sketch of a eukaryote cell

The constituent cells of all but the simplest multicellular animals, both aquatic and terrestrial, exist in an
"internal sea" of extracellular fluid (ECF) enclosed within the integument of the animal from which the
cells take up oxygen and nutrients and into which metabolic waste products are discharged (Ganong.
2005). This harks back to their origins in the primordial seas of the earth, according to the evolution theory
(Cooper, 2000; Ganong, 2005). Because cells originated in a sea of organic molecules, they were able to
obtain food and energy directly from their environment (Ganong, 2005). But such a situation is selflimiting, so cells needed to evolve their own mechanisms for generating energy and synthesising the
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molecules necessary for their replication (Cooper, 2000). The generation and controlled utilisation of
metabolic energy is central to all cell activities, and the principal pathways of energy metabolism are highly
conserved in present-day cells. All cells use adenosine 5-triphosphate (ATP) as their source of metabolic
energy to drive the synthesis of cell constituents and carry out other energy-requiring activities, such as
movement (Guyton and Hall, 2005; Ganong, 2005; Alberts et al, 2002; Cooper, 2000).

The Cell Theory

Ever since the existence of the cell was first demonstrated by Robert Hooke in the seventeenth century, and
with further work done by biologists and later biomedical scientists (starting from the nineteenth century
with the expansion and consolidation of the basic medical sciences in that century), there has evolved a set
of agreed upon propositions about the nature of the cells. These propositions were postulated as the Cell
Theory. The cell theory is a scientific attempt to account for the observed properties of cells (Cooper,
2000). In its simplest enunciated form, it was a triad of statements that describe the origin, form and
function of cells. The cell theory was first formulated in 1838 by Matthias Schleiden and Theodor
Schwann. However, many other scientists like Rudolf Virchow also contributed to the theory. The three
original tenets of the cell theory were propounded as described below:
i. All living organisms are composed of one or more cells. (Viruses, when discovered, were treated
as a special form of life, in many cases being treated more as intelligent molecule than a living
one);
ii. The cell is the basic unit of structure and organisation in organisms;
iii. Cells arise from pre-existing cells;

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The cell theory became the foundation of modern biology and is the most widely accepted explanation of
the function of cells (Neuhaus, 2013; Alberts et al, 2002; Cooper, 2000). As a result of this, the original
propositions have been rendered of near-axiomatic status. A modern and contemporary interpretation of
these propositions, reproduced below (from Neuhaus, 2013), directs and actuates all knowledge, study and
demonstration of biological processes (including in the study of biomedical science, and not to mention
that healthcare interventions depend on insights gleaned from the cell theory).

The generally accepted modern interpretations of the cell theory include:

i. All known living things are made up of one or more cells
ii. All living cells arise from pre-existing cells by division.
iii. The cell is the fundamental unit of structure and function in all living organisms.
iv. The activity of an organism depends on the total activity of the independent cells that constitute it
(coordinated into one smooth industrial flow)
v. Energy flow occurs within cells
vi. Heredity information (DNA) is passed on from cell to cell. Cells contain DNA which is found
specifically in the chromosome and RNA found in the cell nucleus and cytoplasm.
vii. All cells are basically the same in chemical composition in organisms of similar species

The foregoing is deemed necessary background to refresh the mind of just what type of cells make up the
human organism so that there may be a very fruitful discussion of the survival backup system this complex
of cells that is the human organism have evolved for its thriving in incommoding circumstances.

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III
CONSIDERATIONS IN CELL DEVELOPMENT
Introduction
It has been noted earlier in this paper that every cell that makes up the human organism is specialised for a
certain function or a certain group of functions, depending on the structural modifications that it possesses
and which distinguishes it from all other cells (Alberts et al, 2002), and it was noted further that every cell
aggregates with similar cells to function (or just simply exist) as a kind of tissue which may in turn
combine in varying permutations and conjunctions to give the bodys organs and thus on to organ systems
till the whole organism itself is organised (Guyton and Hall, 2005). The point here is that this fact of
organismic organisation is axiomatic of all cell types except for the class of cells already described as stem
cells. These stem cells are properly the primordial parents of each human organism as these are the initial
cells that any individual ever possesses (before individuality is even achieved). Stem cells give rise to other
cells of the body, which now participate in the epigenetic hierarchical organisation that is embryological
development. For a better understanding of the stem cell concept, one must revisit the very beginning of
life.

Embryological Considerations
Human organisms develop from a single cell a fertilised ovum (Moore et al, 2013). The ovum is the
female gamete which fuses with the male gamete in the process of fertilisation to yield the zygote as the
fertilised ovum comes to be known which by a process of rapid mitotic divisions cleavage soon
attains a critical mass (called an embryo) for implantation in the already prepared uterus (which preparation
is part of the cascade of activities referred to as the female ovulation) (Moore et al, 2013; Sadler, 2003).

Embryologists have determined that implantation occurs between eight to twelve days after fertilisation has
occurred i.e. within the first two weeks of embryonic life (Sadler, 2003). Before implantation occurs, each
of the cells (blastomeres) of these cleavage-stage embryos are undifferentiated i.e. are not yet committed to
becoming any particular type of differentiated cell and thus possess the potential to give rise to any cell of
the body (Yu and Thomson, 2006). The first differentiation event in humans occurs at approximately five
days of development, when an outer layer of cells committed to becoming part of the placenta (the
trophectoderm) separates from the inner cell mass (International Society for the Study of Stem Cells, 2016;
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Moore et al, 2013). The cells of the inner cell mass have the potential to generate any cell type of the body,
but after implantation, they are quickly depleted as they differentiate to other cell types with more limited
developmental potential (Moore et al, 2013; Sadler, 2003). However, if these cells are removed from their
normal embryonic environment and cultured under appropriate conditions, they can continue to proliferate
and replicate themselves indefinitely and still maintain the developmental potential to form any cell type of
the body (Yu and Thomson, 2006). These are the embryonic stem cells, and the special completely
undifferentiated capability they possess is termed pluripotency (International Society for the Study of Stem
Cells, 2016; Moore et al, 2013; Yu and Thomson, 2006).

The Cell Reserve and Subsidy System

Not all the differentiated cells become terminally specialised, and these latter type of stem cells retain a
very broad capability of differentiation to the cell types of a distinct tissue/organ. They are thus termed
multipotent stem cells, or alternatively adult/somatic stem cells. These cells are maintained as a store of
cells, as already noted elsewhere in this paper, for the replenishment of highly specialised cells that do not
replicate as well as for the repair of wear and tear in some vital tissues and organs (International Society for
Stem Cell Research, 2016; National Institutes of Health, 2015). Thus, the bodys store of stem cells
functions not unlike a reserve that makes regular subsidies to maintain the continuing viability of the
living human organism (National Institutes of Health, 2015). These stem cell repositories are to be found
in the bone marrow, the gut, and adipose tissue principally, though several other depots exist also in diverse
living human tissue.

The most familiar products of this system are the various components of blood tissue, generated in
erythropoiesis. Less familiar are the cells of the gastrointestinal system, which also have a high turnover
rate due to the very active roles they play in metabolism, the factor that necessitated the existence of a cell
subsidy system to start with. The normal expectation would be that the subsidy system would be activated
every time to replace worn-out or diseased tissue in any part of the body but it does not work out quite like
that, but scientists have been working since the latter part of the last century to investigate ways of
harnessing the enormous potential of these stem cells.

Stem Cells
There are two basic types of stem cells, as already noted the embryonic stem cells and the non-embryonic
(adult or somatic) stem cells (International Society for Stem Cell Research, 2016; National Institutes of
Health, 2015).
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Stem cell differentiation (NIH, 2015)

Stem cells are distinguished from other cell types by two important characteristics. First, they are
unspecialised cells capable of renewing themselves through cell division, sometimes after long periods of
inactivity. Second, under certain physiologic or experimental conditions, they can be induced to become
tissue- or organ-specific cells with special functions. In some organs, such as the gut and bone marrow,
stem cells regularly divide to repair and replace worn out or damaged tissues. In other organs, however,
such as the pancreas and the heart, stem cells only divide under special conditions (National Institutes of
Health, 2015; Yu and Thomson, 2006).

Characteristics of Embryonic Stem Cells (NIH, 2015)

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Embryonic stem cells are unique in their ability to renew themselves indefinitely by producing identical
cells. In addition, under certain physiologic or experimental conditions, they can be induced to give rise to
many different types of cells, each with specialized functions (International Society for Stem Cell
Research, 2016; National Institutes of Health, 2015).

Source of embryonic Stem cell

Non-embryonic stem cells exist in specialised depots in adult human tissue (adult here should be noted to
cover all persons beyond the initial stages of epigenetic development and in whom there is considerable
tertiary specialisation of diverse cell populations) and are important for normal tissue maintenance as well
as for repair after injury. The fact that these cells can be induced proliferate and differentiate to form any
structure in the body, in effect start development locally elsewhere, is what makes them an especial interest
of biomedical research in recent times (International Society for Stem Cell Research, 2016; National
Institutes of Health, 2015). Further, there is also some research progress in deriving a distinct synthetic
analogue of the embryonic stem cell from the somatic stem cell. This is discussed further under in Current
Developments: Stem Cell Research later in this term paper.

We will now turn aside to briefly consider the concomitant focus of this term paper Tissue Culture to
enable a proper web of analysis to be spun. As tissue culture is a biomedical research technique, the
discussion will proceed with a general overview of biomedical research.
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IV
APPROACHES TO BIOMEDICAL STUDY
Introduction
Biomedical research is the broad area of science that is devoted to the study of the processes of life. More
narrowly defined, as given by the New Jersey Association for Biomedical Research (NJABR), biomedical
research is concerned with the investigation of ways to prevent and treat diseases that cause illness and
death in people and in animals as well as to further understand the genetic background to disease and health
and the environmental factors related it (NJABR website, 2016).

This general field of research includes many areas of both the life and physical sciences, and there is a
large amount of cross-disciplinary effort (both multi-disciplinary and inter-disciplinary) deployed in
biomedical research (Sinha and Kumar, 2008; Mathers and Roberts, 1998). This field has seen such rapid
progress in the last thirty to fifty years, with much of the advances being seen only in the last fifteen years
that the whole field of life sciences, pure and applied, has been described as undergoing a paradigmatic
revolution equal to that which occurred in scholarship as whole during the Renaissance and again seen in
science at the dawn of the twentieth century. Already, life and biomedical scholarship has been so radically
altered that many concepts and theories long held sacrosanct are being upturned regularly to be replaced by
newer ones which draw on the emerging new insights.

There are three levels of biomedical study/research delineated by the New Jersey Association for
Biomedical Research, and these were given as Basic (Pure) Research, Applied Research, and Clinical
Research (NJABR, 2016). These categories are dilated upon below.
Basic Research: This is conducted to increase the base knowledge and understanding of the
physical, chemical, and functional mechanisms of life processes and disease. It is fundamental and
not especially directed to solving any particular biomedical problem in humans or animals. This
type of research often involves observing, describing, measuring, and experimental manipulation
and provides the building blocks upon which the other types of research (applied and clinical) are
based. A basic researcher seeks to add to the store of knowledge about how living things work. A
basic researchers experiments add pieces to the immensely complex puzzles of life. The
knowledge gleaned from the conduct of basic research constitutes the core of the biomedical
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science taught to beginners in this field to equip them with necessary knowledge, outlook and
bench skills to understand and interrogate the living organism appropriately
Applied Research: This is rather directed towards specific objectives such as the development of a
new drug, therapy, or surgical procedure. It involves the application of existing knowledge, much of
which is obtained through basic research, to a specific biomedical problem and is termed
translational research. Applied research can be conducted with animals, non-animal alternatives
such as computer models or tissue cultures, or with humans. It is evident that the main focus of
discussion lies primarily in this realm. But first the third category must be touched upon.
Clinical Research: Here, the knowledge gained in basic and applied research is used to conduct
studies (generally on humans) on better and more effective interventions. Studies that take place in
a hospital or clinical setting are focused on the treatment of specific human and animal diseases and
other ailments. Clinical research builds upon the knowledge learned through applied and basic
research. Clinical research is conducted on human beings and takes shape in treatments and drugs
that directly improve human healthcare.
Tissue culture is an important tool for the study of the biology of cells from multicellular organisms as it
provides an in vitro (ex vivo, to be semantically accurate) model of the tissue in a well-defined environment
which can be easily manipulated and analysed. Even though tissue culture has been delineated above as a
procedure of applied research, the three domains of biomedical research identified may, and frequently do,
utilise the techniques in conducting their studies and experiments (Zurlow et al, 1994).

The next section will now focus solely on Tissue Culture, before the narrative is picked up again to enable
a neat dovetailing of the two concepts that form the rationale of this paper.

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V
Tissue Culture

Flasks containing tissue culture growth medium which provides nourishment to growing cells

Introduction
Tissue culture is the growth of tissues or cells separate from the organism. This is typically facilitated via
use of a liquid, semi-solid, or solid growth medium, such as broth or agar (Mather and Roberts, 1998;
Martin, 1994; Zurlo et al, 1994). In modern usage, tissue culture generally refers to the growth of the cells
of a tissue from a multicellular organism in vitro, or more accurately ex vivo (Martin, 1994); essentially a
cell culture (Mather and Roberts, 1998).
The cells to be cultured may be cells isolated from a donor organism (where it is disaggregated directly
from tissue by enzymatic or mechanical means before cultivation), primary cells, or an immortalised cell
line (Mather and Roberts, 1998; Martin, 1994). The cells are cultured by submersion in a biological
substrate, the culture medium, which contains the essential nutrients and energy sources necessary for the
cells' survival (Martin, 1994), development and replication of function, where needed (Sinha and Kumar,
2008; Mather and Roberts, 1998).

The term tissue culture is thus often used interchangeably with cell culture, and the latter is rather more
preferred in contemporary intellectual discourse (Sinha and Kumar, 2008; Martin, 1994), though some
authors still maintain there is a semantic (if no longer practical) distinction (Zurlo et al, 1994).
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Historical Background
The term tissue culture was coined by the American pathologist, Montrose Thomas Burrows (Carrel et al
1911, via Wikipedia website). The literal meaning of tissue culture refers to the culturing of tissue pieces,
i.e. explant culture, and that was the procedure begun crudely as early as 1885 when Wilhelm Roux
removed a section of the medullary plate of an embryonic chicken and maintained it in a warm saline
solution for several days. By this procedure he established the basic principle of tissue culture.

Other early scientific researchers like Ross Granville Harrison, E. Steinhardt, C. Israeli, and R. A. Lambert
contributed greatly to basic knowledge, and the procedure of this biomedical technique. The zoologist R.G.
Harrison, apart from establishing definitely the techniques of tissue culture, may particularly be regarded as
the first to conduct a proto-stem cell experiment when he demonstrated that the growth of frog embryonic
cells that would give rise to nerve cells in a medium of clotted lymph in 1907 (Mather and Roberts, 1998).

Cell/tissue culture techniques were advanced significantly in the 1940s and 1950s to support research in
virology. Growing viruses in cell cultures allowed preparation of purified viruses for the manufacture of
vaccines. For example, the injectable polio vaccine developed by Jonas Salk was one of the first products
mass-produced using cell culture techniques. This vaccine was made possible by the cell culture research
of John Franklin Enders, Thomas Huckle Weller, and Frederick Chapman Robbins, who were awarded a
Nobel Prize for their discovery of a method of growing the virus in monkey kidney cell cultures (Zurlo et
al, 1994).

Basic Concepts and Procedure of Cell/Tissue Culture

i. Isolation of Cells: Cells can be isolated from tissues for culture in several ways. Cells can be easily
purified from blood (however, only the white cells are capable of growth in culture). Mononuclear
cells can be released from soft tissues by enzymatic digestion with enzymes which break down the
extracellular matrix. Alternatively, pieces of tissue can be placed in growth media, and the cells that
grow out are available for culture. This method, as already noted above, is known as explant culture
and is the parent tissue culture technique. Cells cultured directly from a subject are known as primary
cells. With the exception of some derived from tumours, most primary cell cultures have limited
lifespan.
ii. Primary Culture: This refers to the stage of the culture after the cells are isolated from the tissue and
proliferated under the appropriate conditions until they occupy all of the available substrate (i.e.,
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reach confluence). At this stage, the cells have to be subcultured (i.e., passaged) by transferring them
to a new vessel with fresh growth medium to provide more room for continued growth.
iii. Maintaining Cells in Culture: Cells are grown and maintained at an appropriate temperature and
gas mixture (typically, 37 C, 5% CO2 for mammalian cells) in a cell incubator. Culture conditions
vary widely for each cell type, and variation of conditions for a particular cell type can result in
different phenotypes. Majority of isolated primary cells undergo the process of senescence and stop
dividing after a certain number of population doublings while generally retaining their viability (the
Hayflick limit). The most commonly varied factor in culture systems, however, is the cell growth
medium. Recipes for growth media can vary in pH, glucose concentration, growth factors, and the
presence of other nutrients. The growth factors used to supplement media are often derived from the
serum of animal blood, such as fetal bovine serum (FBS), bovine calf serum, equine serum, and
porcine serum.
iv. Cell Lines: After the first subculture, the primary culture becomes known as a cell line or subclone.
Cell lines derived from primary cultures have a limited life span (i.e., they are finite; see below), and
as they are passaged, cells with the highest growth capacity predominate, resulting in a degree of
genotypic and phenotypic uniformity in the population.
v. Cell Strain: If a subpopulation of a cell line is positively selected from the culture by cloning or
some other method, this cell line becomes a cell strain. A cell strain often acquires additional genetic
changes subsequent to the initiation of the parent line.
vi. Manipulation of Cultured Cells: The common manipulations carried out on culture cells are media
changes, passaging cells, and transfecting cells. These are generally performed using tissue culture
methods that rely on aseptic technique. Aseptic technique aims to avoid contamination with bacteria,
yeast, or other cell lines. Antiobiotics are usually added to ensure no microbial contamination of the
culture (and the first real advances over the crude early techniques was seen only when the first
antibiotic drugs became available in the 1940s). As cells undergo metabolic processes, acid is
produced and the pH decreases. Often, a pH indicator is added to the medium to measure nutrient
depletion.
vii. Cryopreservation: If a surplus of cells is available from subculturing, they are usually treated with
an appropriate protective agent (e.g., DMSO or glycerol) and stored at temperatures below 130C
(cryopreservation) until they are needed.

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Culture Conditions
Culture conditions vary widely for each cell type, but the artificial environment in which the cells are
cultured invariably consists of a suitable vessel containing the following:

Substrate or medium that supplies the essential nutrients (amino acids, carbohydrates, vitamins,
minerals)

Growth factors

Hormones

Gases (O2, CO2)

Regulated physico-chemical environment (pH, osmotic pressure, temperature)

Most cells are anchorage-dependent and must be cultured while attached to a solid or semi-solid substrate
(adherent or monolayer culture), while others can be grown floating in the culture medium (suspension
culture).

Morphology of Cells in Culture

Cells in culture can be divided into three basic categories based on their shape and appearance in the
culture:
Fibroblast-like cells,

Bipolar or multipolar

Have elongated shapes

Grow attached to a substrate

Epithelial-like cells,

Polygonal in shape with more regular dimensions

Grow attached to a substrate in discrete patches.

Lymphoblast-like cells,

Spherical in shape

Usually grown in suspension without attaching to a surface.

Other types exist, however, the most common usually being the neuronal cell (Martin, 1994).

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Technical Issues in Culturing

As cells generally continue to divide in culture, they generally grow to fill the available area or volume.
This can generate several issues, several of which have been classical issues which were only slowly
overcome (Sinha and Kumar, 2008; Mather and Roberts, 1998; Martin, 1994; Zurlow et al, 1994):

Nutrient depletion in the growth media

Changes in pH of the growth media

Accumulation of apoptotic/necrotic (dead) cells

Cell-to-cell contact, which can stimulate cell cycle arrest, causing cells to stop dividing, known as
contact inhibition. Cell-to-cell contact can also stimulate cellular differentiation.

Recent advances have thrown up another technical issue genetic and epigenetic alterations. This is
usually accompanied by a natural selection of the altered cells potentially leading to overgrowth of
abnormal, culture-adapted cells with decreased differentiation and increased proliferative capacity (Nguyen
et al, 2012).
Other Issues and Current Concepts in Contemporary Cell Culture
i. Finite versus Continuous Cell Line: Normal cells usually divide only a limited number of times
before losing their ability to proliferate, which is a genetically determined event known as
senescence. These cell lines are known as finite. However, some cell lines become immortal through
a process called transformation, which can occur spontaneously or can be chemically or virally
induced.

When a finite cell line undergoes transformation and acquires the ability to divide

indefinitely, it becomes a continuous cell line.

ii. Cell line Cross Contamination: This can be a problem for scientists working with cultured cells.
Such contamination poses a problem for the quality of research produced using cell culture lines, and
the major repositories are now authenticating all cell line submissions. To address this problem of cell
line cross-contamination, researchers are encouraged to authenticate their cell lines at an early
passage to establish the identity of the cell line, and to repeat authentication before freezing cell line
stocks every two months during active culturing and before any publication of research data
generated using the cell lines. Many methods are used to identify cell lines, including isoenzyme
analysis, human lymphocyte antigen (HLA) typing, chromosomal analysis, karyotyping, morphology
and STR analysis
iii.

Established human cell lines: It is possible to fuse normal cells with an immortalised cell line. This
method is used to produce monoclonal antibodies. In brief, lymphocytes isolated from the spleen (or

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possibly blood) of an immunised animal are combined with an immortal myeloma cell line (B cell
lineage) to produce a hybridoma which has the antibody specificity of the primary lymphocyte and
the immortality of the myeloma. Selective growth medium (HA or HAT) is used to select against
unfused myeloma cells; primary lymphoctyes die quickly in culture and only the fused cells survive.
These are screened for production of the required antibody, generally in pools to start with and then
after single cloning. But when this is done with human cells, it gives rise to a host of bioethical
concerns as they may outlive their parent organism and later be used in the discovery of lucrative
medical treatments.
iv.

Two-Dimensional and Three-Dimensional Culture: Traditional tissue culture procedure primarily

involves cultures of cells on flat plastic dishes (Mather and Sinha and Kumar, 2008; Roberts, 1998;
Martin, 1994; Zurlo et al, 1994) a procedure known as two-dimensional (2D) cell culture, first
developed by Wilhelm Roux in 1985 (Sinha and Kumar, 2008; Martin, 1994). Advances in polymer
technology gave rise to plastic Petri dish (named for Jules Petri) which is the standard for
contemporary 2D cell culture but various researchers today also utilise culturing laboratory flasks,
conicals, and even disposable bags like those used in single-use bioreactors (Sinha and Kumar, 2008).
Aside from Petri dishes, however, biomedical scientists have long been growing cells within
biologically derived matrices such as collagen or fibrin (Sinha and Kumar), and more recently, on
synthetic hydrogels such as polyacrylamide or PEG. This is done in order to elicit phenotypes that are
not expressed on conventionally rigid substrates. Studies that investigated the control of matrix
stiffness led to discoveries in fields such as stem cell self-renewal, lineage specification, cancer cell
phenotype, mechano-sensing etc. (Dupont et al, 2011; Gilbert et al, 2010; Levental et al, 2009; Engler
et al, 2006; Discher et al, 2005). While bio-derived matrices such as collagen may be conceptualised
as extended 2D culture procedures, the newer synthetic hydrogels are known as three-dimensional
cell cultures from the effort made to mimic the cells natural environment in vivo the extracellular
matrix. Apart from the hydrogel matrix, [42] other platforms used to support 3D cell cultures are
solid scaffolds, and scaffold-free systems such as low-adhesion plates, nanoparticle facilitated
magnetic levitation,[43] and hanging drop plates.[44] As the natural extracellular matrix is important
in the survival, proliferation, differentiation and migration of the cells, different hydrogel culture
matrices mimicking the natural extracellular structure are considered as potential approaches towards
in vivo like cell culturing.

v.

Tissue Engineering: Cell culture is a fundamental component of tissue engineering, as it establishes

the basics of growing and maintaining cells in vitro. Tissue engineering is possible because cells will
normally aggregate where there is an effective extracellular matrix to support that kind of
development. Advances recorded in culture techniques specifically 3D culture, as well as progress

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in newer fields such as bioinformatics e.g. distributed intelligence and 3D printing have
revolutionised the whole concept of tissue engineering that were not even conceivable only a few
decades ago. Tissue engineering potentially offers dramatic improvements in low cost medical care
for hundreds of thousands of patients annually.
vi.

Alternative Cell Culture: Most complex biological products of cell culture are produced by
recombinant DNA (rDNA) technology in animal cell cultures due to the necessary glycosylation
carbohydrate-modified needed in such molecule, and which can currently only be made in animal
cells e.g. the hormone erythropoietin. The cost of growing mammalian cell cultures is high, so
research is underway to produce such complex proteins in insect cells or in higher plants. The use of
single embryonic cell and somatic embryos as a source for direct gene transfer via particle
bombardment are also being investigated, and transit gene expression and confocal microscopy
observation are some of the early applications of this new technique. It also offers to confirm single
cell origin of somatic embryos and the asymmetry of the first cell division, which starts the process
(Duell et al, 2011).

Applications of Cell Culture

i.

Cell culture is one of the major tools used in cellular and molecular biology research, providing
excellent model systems for studying the normal physiology and biochemistry of cells (e.g.,
metabolic studies, ageing), the effects of drugs and toxic compounds on the cells, and mutagenesis
and carcinogenesis.

ii.

It is also used in drug screening and development, and large scale manufacturing of biological
compounds e.g., vaccines, therapeutic proteins (). Mass culture of animal cell lines is particularly
fundamental to the manufacture of viral vaccines and other products of biotechnology (). The major
advantage of using cell culture for any of these applications is the consistency and reproducibility
of results that can be obtained from using a batch of clonal cells.

iii.

The major application of human cell culture is in stem cell industry. Culture of human stem cells is
used to expand the number of cells and differentiate the cells into various somatic cell types for
transplantation (Quiana and Saltzmann, 2004). Stem cell culture is also used to harvest the
molecules and exosomes that the stem cells release for the purposes of therapeutic development
(Maguire, 2016). Mesenchymal stem cells can also, and are, cultured and cryopreserved for future
use.

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VI
STEM CELL RESEARCH

( N I H, 2 0 15 )

The term stem cell was coined towards the end of the nineteenth century to describe certain cells of the body that
were discovered to have the ability to develop into more than one type of body cell (National Institutes of Health,
2015). The first stem cells to be identified were the haematopoietic stem cells of the bone marrow, and efforts
commenced almost immediately to apply the new knowledge to therapeutic intervention (International Society for
Stem Cell Research, 2016; National Institutes of Health, 2015). Limited successes were recorded however, and
further advances had to wait for further advances in biomedical research techniques and immunological knowledge
(which field was born around the same time as knowledge of stem cells began to develop) (International Society for
Stem Cell Research, 2016; Yu and Thomson, 2006).

Historical Progress
The latter half of the twentieth century saw giant strides being made in biomedical knowledge and research
techniques saw increasing breakthroughs being made in stem cell research. Particularly, the explosion in scientific
knowledge (much of it emanating from the unethical experiments conducted in Nazi Germany) facilitated these
developments and by the early 1950s bone marrow transplants were already being successfully done, and this
perhaps represents the first real fruit of what came to be later known as stem cell research. Efforts continued through
the 1960s and 1970s, but stem cell research continued to be conducted under the auspices of the traditional
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biomedical fields their associated clinical specialties e.g. haematology. The field itself was born only after the first
stem cell was successfully isolated. (International Society for Stem Cell Research, 2016; National Institutions of
Health, 2015; Yu and Thomson, 2006)

The year 1981 marks the initial milestone of modern stem cell research as this was the year the first stem
cells were successfully isolated from mice (Martin, 1981; Evans and Kaufman, 1981) but detailed study of
the biology of mouse stem cells did not yield discovery of a method to derive stem cells from human
embryos human embryonic stem cells and grow the cells in the laboratory until 1998 (Yu and
Thomson, 2006) immediately.

Although attempts to derive human embryonic stem cells were made as early as the 1980s, culture media
for human embryos produced by in vitro fertilisation were suboptimal (the embryos used in these studies
were created for reproductive purposes through in vitro fertilisation procedures, and when they were no
longer needed for that purpose, they were donated for research with the informed consent of the donor).
Thus, it was difficult to culture single-cell fertilized embryos long enough to obtain healthy blastocysts for
the derivation of embryonic stem cell lines. Also, species-specific differences between mice and humans
meant that experience with mouse embryonic stem cells was not completely applicable to the derivation of
human embryonic stem cells (Yu and Thomson, 2006). However, in the 1990s, embryonic stem cell lines
from two non-human primates, the rhesus monkey and the common marmoset, were derived, and these
offered closer models for the derivation of human embryonic stem cells. Experience with non-human
primate embryonic stem cell lines and improvements in culture medium for human in vitro fertilisationproduced embryos led rapidly to the derivation of human embryonic stem cell lines at last in 1998 (Yu and
Thomson, 2006).

Current Developments
Until recently, scientists worked with both kinds of stem cells but traditional research for interventional
application has always been focused on embryonic stem cells because of its pluripotential nature
(International Society for Stem Cell Research, 2016; National Institutes of Health, 2015; Yu and Thomson,
2006), though most of the actual interventional breakthroughs made have come from working with somatic
stem cells (). The embryonic stem cells always had to be harvested from discarded embryos as a by-product
of fertility procedures. Another source of embryonic stem cells was been developed in the last fifteen years,
and it is the use of a technique called somatic cell nuclear transfer, also referred to a therapeutic cloning.

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In this technique, nuclei from the cells of living patients with specific diseases are isolated and used for the
generation of embryonic stem cells. This is achieved by placing one such nucleus into a donated
unfertilized egg from which the genetic material has been removed and then stimulating the egg to divide
to the stage when stem cells can be derived in culture. These cells can then be induced to develop in the
laboratory into specialized cells such as nerve cells that are affected by the disease in question. In the
future, research can elucidate the pathogenesis of different diseases and determine the best interventions
that may be used to treat it, just by laboratory observation of these cells to discover how they differ from
normal cells over time. In addition, embryonic stem cells derived by this technique ultimately may be
useful for cell-based replacement therapy for the original patients from whom they were derived. This is
because they are genetically identical to the patient and are not rejected by the immune system (Wilmut and
Paterson, 2003). While somatic nuclear transfer is an important step toward realizing the promise of stem
cell research, the approach raises new ethical and regulatory issues that must be addressed carefully
(Magnus and Cho, 2006).

It is, however, being considered that non-embryonic stem cells may ultimately become useful as
alternatives to embryonic stem cells in therapeutic, and investigation into the therapeutic potential of adult
stem cells has been on for some time (Weiss et al, 2006; American Thoracic Society, 2006), and some
preliminary success is being declared, albeit cautiously (International Society for Stem Cell Research,
2016). About ten years ago, researchers made a breakthrough by identifying conditions that would allow
some specialised adult cells e.g. skin cells to be "reprogrammed" genetically to assume a stem cell-like
state. This new type of stem cell is known as the induced pluripotent stem cell (iPSC), and this is already
being presented as a third type of stem cell but this usage lacks widespread acceptance as at yet
(International Society for Stem Cell Research, 2016; National Institutes of Health, 2015).

An induced pluripotent stem cell (NIH, 2015)

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Proponents of making it a third category of stem cells point out, quite rightly, that while induced
pluripotent stem cells share many of the same characteristics of embryonic stem cells, including the ability
to give rise to all the cell types in the body, induced pluripotent stem cells and embryonic stem cells are in
no way identical to each other. The more conservative prefer to view them at the moment as extremely
diversified hybridised derivatives of somatic stem cells, least of all not bing that these cells do not occur in
nature. The original induced pluripotent stem cells were produced by using viruses to insert extra copies of
three to four genes known to be important in embryonic stem cells into the specialized cell. It is not yet
completely understood how these three to four reprogramming genes are able to induce pluripotency, and
this question (amongst many others) is the focus of ongoing research.

Multiple studies are still investigating the various ramifications of the induced pluripotent stem cells, and it
is easily the hottest topic in stem cell research at the present time. One of the major advantages of induced
pluripotent stem cells, ( which is one of the reasons of their current popularity in research), is that they are
a very good way to make pluripotent stem cell lines that are specific to a disease or even to an individual
patient. Disease-specific stem cells are powerful tools for studying the cause of a particular disease and
then for testing drugs or discovering other approaches to treat or cure that disease. The development of
patient-specific stem cells is also very attractive for cell therapy, as these cell lines are from the patient
themselves and may minimise some of the serious complications of rejection and immunosuppression that
can occur following traditional donor-facilitated tissue and organ transplant surgeries.

Clinical Applications
Translation: Clinical translation is the process used to turn basic scientific knowledge into real-world
medical treatments. Researchers use new insights into the normal morphology and physiology a tissue
usually, and the pathology and pathogenesis of particular diseases or injuries that may occur in it and use
this information to develop new diagnostic interventional procedures. Before being marketed or adopted as
standard of care, however, most interventions are tested through clinical trials. Nonetheless, sometimes, in
attempting new surgical techniques or where the disease or condition is rare and does not have a large
enough population to form a clinical trial, certain treatments might be tried on one or two people ( a form
of testing sometimes referred to as innovative medicine). These two approaches to translational research
have been applied, and are being used currently, in making available clinically the insights and progresses
achieved in stem cell research.

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Current Therapies: Blood stem cells are currently the most frequently used stem cells for therapy. For
more than fifty years, physicians have been using bone marrow transplants to transfer blood stem cells to
patients, and more advanced techniques for collecting blood stem cells are now being used to treat
leukaemia, lymphoma and several other blood disorders. Umbilical cord blood, like bone marrow, is often
collected as a source of blood stem cells and in certain cases is being used as an alternative to bone marrow
transplantation. Additionally, some bone, skin and corneal diseases or injuries can be treated by grafting
tissues that are derived from or maintained by stem cells. These therapies have also been shown to be safe
and effective.
Potential Therapies: Other stem cell treatments, while promising, are still at very early experimental
stages. For example, the mesenchymal stem cell, found throughout the body including in the bone marrow,
can be directed to become bone, cartilage, fat and possibly even muscle (International Society for Stem
Cell Research, 2016). In certain experimental models, these cells also have some ability to modify immune
functions. These abilities have created considerable interest in developing ways of using mesenchymal
stem cells to treat a range of musculoskeletal abnormalities, cardiac disease and some immune
abnormalities such as graft-versus-host disease following bone marrow transplant (Jung and Kleinheinz,
2014).

Challenges: Despite the recorded successes, and the attractive promise, of stem cell research, it is
apparent that there are several major challenges that must be addressed before its full potential to study cell
processes and its utility as cell therapies to treat a wider range of diseases can be fully exploited.
1. Source of cells. There is a need to identify an abundant source of stem cells. The identification,
isolation and culture of the right kind of stem cell, particularly in the case of rare adult stem cells,
are painstaking and difficult processes. Pluripotent stem cells, such as embryonic stem cells, can be
grown indefinitely in the lab and have the advantage of having the potential to become any cell in
the body, but these processes are again very complex and must be tightly controlled. Induced
pluripotent stem cells, while promising, are also limited by these concerns. In both cases,
considerable work remains to be done to ensure that these cells can be isolated and used safely and
routinely.
2. Tissue match. As with organ transplants, it is very important to have a close match between the
donor tissue and the recipient; the more closely the tissue matches the recipient, the lower the risk
of rejection. Being able to avoid the life-long use of immuno-suppressants would also be preferable.
The discovery of induced pluripotent stem cells has opened the door to developing patient-specific

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pluripotent stem cell lines that can later be developed into a needed cell type without the problems
of rejection and immunosuppression that occur from transplants from unrelated donors.
3. Product Delivery. A system for delivering the cells to the right part of the body must be developed.
Once in the right location, the new cells must then be encouraged to integrate and function together
with the bodys other cells. This means that advances in immunology, especially those with
biotechnological applications, must keep pace with other developments in the biomedical arena if
the insights gleaned in other areas will be clinically translatable.

Ethical Considerations
If the outcome of a certain research were known in advance, the research would not be necessary in the
first place. And the benefits of research, like its results, cannot be completely specified in advance.
Similarly, its costs can never be completely accurately foreseen. Such costs of course include ethical and
social costs. Ethical and social issues arise to some extent whenever scientific research is carried out,
because the outcome affects people. In particular, such issues arise in biomedical research because the
interests of potential beneficiaries may compete with, and may have to be considered together with costs to
society or to other individuals, such as donors. Stem cell research includes theoretical (or basic) and
applied (or practical) aspects. The main intended benefits are:
Theoretically, advancing understanding of tissue differentiation, development, repair and ageing.
Practically, the therapeutic use of undifferentiated tissue for organ/tissue replacement or repair.
The distinction between theoretical and applied research, in any field, is one of time scale. In the long run,
theoretical advances find application. In the short term, research can address immediate problems. Others
problems may arise unanticipated, however. Stem cell research, especially embryonic stem cell research is
generally held to pose a classic moral dilemma:

The duty to prevent or alleviate suffering

The duty to respect the value of human life

It is generally agreed that in the case of embryonic stem cell research, it is impossible to respect both moral
principles. To obtain embryonic stem cells, the early embryo has to be sacrificed. This has been interpreted
by critics of stem cell research to mean destroying a potential human life (). Its proponents, on the other
hand, contend that embryonic stem cell research could lead to the discovery of new medical treatments that
would alleviate the suffering of many people.
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This has engendered, both in scientific circles and policy-making conclaves, an intractable conundrum
attended by an acrimoniously polemical debate. The solution to the debate hinges on the view adopted of
the embryo. Basically, does it (an embryo) have the moral status of a person, or not? If it does, then it has
rights that must be respected and protected. If it does not, well then critics and opponents have only making
a mountain out of a molehill. The whole issue is confused further by the fact that the contenders are not so
much interested in finding out where the truth lies but only in justifying their own preconceived notions
(which are often derived from some unscientific premises while yet purporting to adduce arguments based
on scientific knowledge and insights).

Kristina Hug (2015) presented four fundamental polemical views of the moral status of the embryo. These
views move from a pole of Sanctity to a pole Utility, traversing two Middle-of-the-Road Views in the
process. These views encapsulate the major arguments for and against stem cell research based on the
origin of the stem cell. Research involving somatic stem cells does not normally invite this category of
objections though the recent induced pluripotent stem cell technique invites similar debate, but to greater or
lesser extents depending on the exact research protocol followed. The views are proposed are presented
below:
1st View: The embryo has full moral status from fertilisation onwards
The polemic is strongest and sharpest here. Either the embryo is viewed as a person whilst it is still an
embryo, or it is seen as only a potential person. Complicating issues is that the criteria for personhood
are notoriously unclear, and different people define what makes a person in different ways.
Arguments for this view
Development from a fertilized egg into to
baby is a continuous process and any
attempt to pinpoint when personhood
begins is arbitrary.
A human embryo is a human being in the
embryonic stage, just as an infant is a
human being in the infant stage.
Although an embryo does
not currently have the characteristics of a
person, it will become a person and should
be given the respect and dignity of a
person.

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Arguments against this view

An early embryo that has not yet implanted into the uterus
does not have the psychological, emotional or physical
properties that we associate with being a person. It
therefore does not have any interests to be protected and
we can use it for the benefit of patients (who ARE
persons).
The embryo cannot develop into a child without being
transferred to a womans uterus. It needs external help to
develop. Even then, the probability that embryos used for
in vitro fertilization will develop into full-term successful
births is low. Something that could potentially become a
person should not be treated as if it actually were a person

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This is the classic or the All-or-None debate. It is also regarded as the Sanctity View, as it believes in
the complete sanctity of the embryo as human life.
2nd View: There is a cut-off point at 14 days after fertilization
This represents an attempt to find a middle way between the two opposing viewpoints (1st, or Fixed,
Middle-of-the-Road View). It proposes special protection for the human embryo only from after day 14
after fertilization because:

After 14 days the embryo can no longer split to form twins. Before this point, the embryo could still
be split to become two or more babies, or it might fail to develop at all.

Before day 14, the embryo has no central nervous system and therefore no senses. If we can take
organs from patients who have been declared brain dead and use them for transplants, then we can
also use hundred-cell embryos that have no nervous system.

Fertilization is itself a process, not a moment. An embryo in the earliest stages is not clearly
defined as an individual.

3rd View: The embryo has increasing status as it develops

This is also a Middle-of-the-Road view, but leaning heavily to the pole of Utility (4th view) as opposed
to Sanctity (1st view). It agrees with the preceding two views that an embryo deserves some protection
from the moment the sperm fertilizes the egg. However, this view holds that the level of protection
afforded an embryo should increase only its moral status increases i.e. as it becomes more human-like.
There is thus no homogeneity amongst the proponents of this view and they span the spectrum of
difference between
Arguments for this view
Arguments against this view
There are several stages of development We protect a persons life and interests not because
that could be given increasing moral
they are valuable from the point of view of the
status:
universe, but because they are important to the
i. Implantation of the embryo into the
person concerned. Whatever moral status the human
uterus wall around six days after
embryo has for us, the life that it lives has a value to
fertilization.
the embryo itself.
ii. Appearance of the primitive streak If we judge the moral status of the embryo from its
the beginnings of the nervous
age, then we are making arbitrary decisions about
system at around 14 days.
who is human. For example, even if we say
iii. The phase when the baby could
formation of the nervous system marks the start of
survive if born prematurely.
personhood, we still would not say a patient who has
iv. Birth.
lost nerve cells in a stroke has become less human.
If we are not sure whether a fertilized egg should be
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 If a life is lost, we tend to feel differently

about it depending on the stage of the
lost life. A fertilized egg before
implantation in the uterus could be
granted a lesser degree of respect than a
human fetus or a born baby.
More than half of all fertilized eggs are
lost due to natural causes. If the natural
process involves such loss, then using
some embryos in stem cell research
should not worry us either.

considered a human being, then we should not

destroy it. A hunter does not shoot if he is not sure
whether his target is a deer or a man.

4th View: The embryo has no moral status at all

This view is that An embryo is organic material with a status no different from other body parts.
Arguments for this view
Fertilized human eggs are just parts of other
peoples bodies until they have developed enough
to survive independently.
The only respect due to blastocysts is the respect
that should be shown to other peoples property.
If we destroy a blastocyst before implantation into
the uterus we do not harm it because it has no
beliefs, desires, expectations, aims or purposes to
be harmed.

Arguments against this view

By taking embryonic stem cells out of an
early embryo, we prevent the embryo from
developing in its normal way. This means it
is prevented from becoming what it was
programmed to become a human being.

The most contentious and acrimonious debates of all rages between the 1st and the 4th views of all since it
ranges most theistic believers, especially Christians from mainstream creedal confession (who generally
hold this view) against militant atheistic scientists (who generally hold the fourth view), and the ensuing
debate is many times used as a stalking horse for wider metaphysical disputes.

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VII
REFERENCES
Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., Walter, P. (2002): Molecular Biology of the Cell: A
Problems Approach, 4th ed. Accessed online at http://www.ncbi.nlm.nih.gov/books/NBK26869/
American Thoracic Society (2006): Human Embryonic Stem Cell Research. American Journal of Respiratory and
Critical Care Medicine; 173 (9): 1043-1045
Carrel, Alexis and Montrose T. Burrows: (1911): Cultivation of Tissues in Vitro and its Technique. Journal of
Experimental Medicine; 13: 387-396. Via: https://en.wikipedia.org/wiki/Tissue_culture. Accessed on 30 April
at 0705hrs
Cooper, G.M. (2000): The Cell: A Molecular Approach, 2nd ed. Sunderland: Sinauer Associates. Accessed online at
http://www.ncbi.nlm.nih.gov/books/NBK9925/
Discher, D. E., Janmey, P., Wang, Y.L. (2005): Tissue Cells Feel and Respond to the Stiffness of Their Substrate.
Science; 310(5751): 113943.
Duell, B.L., Cripps, A.W., Schembri, M.A., Ulett, G.C. (2011): Epithelial cell co-culture models for studying
infectious diseases: Benefits and limitations. Journal of biomedicine & biotechnology; 85: 241-249
Dupont, S., Morsut, L., Aragona, M., Enzo, E., Giulitti, S., Cordenonsi, M., Zanconato, F., Le Digabel, J., Forcato,
M., Bicciato, S., Elvassore, N., Piccolo, S. (2011): Role of YAP/TAZ in mechanotransduction. Nature; 474
(7350): 17983.
Engler, A.J., Sen, S., Sweeney, H.L., Discher, D.E. (2006): Matrix Elasticity Directs Stem Cell Lineage
Specification. Cell; 126 (4): 67789
Evans, M.J., Kaufman, M.H. (1981): Establishment in culture of pluripotential cells from mouse embryos. Nature;
292:154156.
Ganong, W.F. (2005): Review of Medical Physiology, 22ed. New York: McGraw Hill Medical.
Gilbert, P.M., Havenstrite, K.L. Magnusson, K.E., Sacco, A., Leonardi, N.A., Kraft, P., Nguyen, N.K., Thrun, S.,
Lutolf, M.P., Blau, H.M. (2010): Substrate elasticity regulates skeletal muscle stem cell self-renewal in culture.
Science; 329 (5995): 107881.
Guyton, A.C., Hall, J.E. (2005): Textbook of Medical Physiology, 11th ed. Philadelphia: Saunders.

29 | P a g e

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137868

International

Society

for

Stem

Cell

Research

(2016):

Stem

Cell

Facts.

Retrieved

from

http://www.isscr.org/docs/default-source/isscr-publications/stem-cell-factsbrochure92203E27C59B.pdf?sfvrsn=2 on 30 April at 0715hrs

Jung, S., Johannes Kleinheinz, J. (2014): Adult Mesenchymal Stem Cells in Current Tissue Engineering Concepts In:
Eberli,

D.

(ed):

Cells

and

Biomaterials

in

Regenerative

Medicine.

Retrieved

from:

http://www.intechopen.com/books/cells-and-biomaterials-in-regenerative-medicine/adult-mesenchymal-stemcells-in-current-tissue-engineering-concepts on 30 April at 0745hrs

Kumar, V., Abbas, A.K., Fausto, N. (2005): Robbins and Cotrans Pathologic Basis of Disease, 7th ed. Philadelphia:
Elsevier Saunders.
Levental, K.R., Yu, H., Kass, L., Lakins, J.N., Egeblad, M.; Erler, J.T.., Fong, S.F.T., Csiszar, K., Giaccia, A.,
Weninger, W., Yamauchi, M., Gasser, D.L., Weaver, V.M. (2009): Matrix Crosslinking Forces Tumor
Progression by Enhancing Integrin Signaling. Cell; 139 (5): 891906.
Maguire, G. (2016) Therapeutics from Adult Stem Cells and the Hype Curve. ACS Medical Chemistry Letters; 7 (5):
441443
Magnus, D., Cho, M.K. (2005): Ethics: issues in oocyte donation for stem cell research. Science; 308:17471748
Mather, J.P., Roberts, P.E. (1998): Introduction to Cell and Tissue Culture: Theory and Technique. New York:
Plenum Press. Accessed at https://www.scribd.com/doc/427679/Introduction-to-Cell-and-Tissue-Culture on
30 April at 0815hrs
Martin, B.M. (1994): Tissue Culture Techniques: An Introduction. Boston: Birkhuser
Martin, G.R. (1981): Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned
by teratocarcinoma stem cells. Proceedings of the National Academy of Science USA; 78:76347638.
Moore, K.L., Persaud, T.V.N., Torchia, M.G. (2013): The Developing Human: Clinically Oriented Embryology, 9th
ed. Philadelphia: Elsevier Saunders.
National Institutes of Health (2015): Stem Cell Information [World Wide Web site]. Bethesda, MD: National
Institutes

of

Health,

U.S.

Department

of

Health

and

Human

Services.

Retrieved

from

http://stemcells.nih.gov/info/basics/pages/default.aspx on 30 April at 0610hrs

Neuhaus, G. (2013): The Structure and Ultrastructure of the Cell. Berlin: Springer.
New Jersey Association of Biomedical Research (2016): Biomedical Research Definitions in: Education; General
Background to Biomedical Research. Retrieved from http://njabr.com/education/general-background-onbiomedical-research/biomedical-research-definitions/ on 1 May, 2016 at 1930hrs
30 | P a g e

Current Concepts and Utility of

Stem Cells and Tissue Culture Research

Ayanleke, E.O.
137868

Nguyen, H. T., Geens, M., Spits, C. (2012). Genetic and epigenetic instability in human pluripotent stem cells.
Human Reproduction Update; 19(2): 187205.
Qiana, L., Saltzman, W.M. (2004) Improving the expansion and neuronal differentiation of mesenchymal stem cells
through culture surface modification. Biomaterials; 25: 1331-1337.
Sadler, T.W. (2003): Langmans Medical Embyology, 9th ed. Philadelphia: Elsevier.
Sinha, K.B., Kumar, R. (2008): Principles of Animal Cell Culture: A Students Compendium. Lucknow: International
Books Publishing Co. Accessed at https://www.scribd.com/doc/37108650/Principles-of-Animal-Cell-Culture
on 1 May, 2016 at 0900hrs
Weiss, D.J., Berberich, A.M., Borok, Z., Gail, D., Kolls, J.K., Penland, C., Prockop, D.J. (2006): National Heart
Lung Blood Institute/Cystic Fibrosis Foundation Workshop Report: Adult stem cells, lung biology, and lung
disease. Proceedings of the American Thoracic Society; 3:193207
Weiss, A.M., Breitenbach, M., Rinnerthaler, M., Virt, G. (2013): Ethical Considerations on Stem Cell Research In:
Bhartiya D. (Ed.): Pluripotent Stem Cells, Accessed at http://www.intechopen.com/books/pluripotent-stemcells/ethical-considerations-on-stem-cell-research on 1 May, 2016 at 0850hrs
Wilmut, I., Paterson, L. (2003): Somatic cell nuclear transfer. Oncological Research; 13:303307.
Yu, J., Thomson, J.A. (2006): Embryonic Stem Cells. In: Stem Cell Information [World Wide Web site]. Bethesda:
National Institutes of Health, U.S. Department of Health and Human Services, 2014. Available at
http://stemcells.nih.gov/info/scireport/Pages/2006Chapter1.aspx. Accessed on 1 May, 2016 at 1900hrs
Zurlo, J., Rudacille, D., Goldberg, A.M. (1994): Animals and Alternatives in Testing: History, Science, and Ethics.
New

York:

Mary

Ann

Liebert,

Inc.

Accessed

at

https://web.archive.org/web/20060210014148/http://caat.jhsph.edu/pubs/animal_alts/glossary.htm on 1 May,
2016 at 1900hrs

31 | P a g e

Current Concepts and Utility of

Stem Cells and Tissue Culture Research

Ayanleke, E.O.
137868