MCB38 Lecture #2: Firestone

Asymmetric Division of
stem cells

Self Differentiation
renewal pathway

Stem Cell Partial Differentiation;

Lineage specific
progenitor cell
stem cells can
differentiated into ~200
different cell types
depending on the
source of the stem cells

Differentiated
1
cell
 Functional relationships of molecules, cells,
tissues, organs and organ systems
that are critical for the normal existence of multicellular organisms
Functions of Molecules
Molecules (DNA, RNA, proteins, etc.)

Gene Therapy Reprogramming:

Gene Expression
Gene Editing
Differentiated
Stem Cells
Cells Hormonal and
Environmental Signals

Tissues Tissue Microenvironment: Cell-Cell

interactions; Cell shape;
Intercellular communication

Organs/organ Multi-tissue organization

systems and functional coordination

Dysfunction and loss of coordination can trigger diseases (such

as diabetes, cancer, etc.) that in theory can be corrected using
stem cell technologies

Cells: Approximately 200 different cell types in a human (~50 trillion cells total)
Tissues: Groups of cells with common functions and structures
Organs: Combination of tissues that forms a functional unit
Organ Systems: Group of organs that work together in the organism
(such as your digestive or reproductive systems)

2
Cells: Stem cells vs Differentiated cells – Gene expression; Specific functions,
Tissue formation; Responsiveness to extracellular stimuli, etc.

Many structural, genetic and biochemical similarities (such as organelles with

specific functions).

“Stemness” gene expression:

1) Expressed in stem cells and either not expressed or expressed at only very low
levels in differentiated cells. (Embryonic stems cells do not express differentiation-
associated genes; although certain adult stem cells can express differentiation-
associated genes). Many “stemness” genes are transcription factors (TF) that bind
to promoters of target genes to stimulate mRNA synthesis.

mRNA Protein Activity

TF

Target gene promoter

2) Expression at high levels in differentiated cells can induce stem cell-like properties
3) Loss of expression in stem cells results in loss of certain “stemness” properties

Tissues:

•  Various structures and shapes depending on function

•  Cells within a tissue can be connected by their extracellular matrix (can be viewed as
weaved together in a fabric of fibers) The term tissue is derived from the Latin word
meaning “weave”

•  Four main categories of tissues

Epithelial tissue
Connective tissue
Nervous tissue
Muscle tissue

3
 Epithelial Tissue
Epithelial cells
Basement membrane

Epithelial Tissue
•  Sheets of tightly packed cells that covers the outside of your body (skin) and
lines the organs and cavities within the body
•  Free surface of epithelial tissue exposed to the environment (air or fluid), and cells
at the base of the epithelial layer are attached to a basement membrane
• Functions as a barrier protecting against injury, invasive microorganisms and fluid loss
•  Some epithelial cells secrete proteins, and absorb nutrients (for example: small intestine)
•  Most cancers are derived from epithelial cells
•  Relatively little material between cells, which are held together by intercellular junctions

Classify epithelia tissue by number of cell layers and shape of cells

Cell Layers: Simple epithelium-single layer of cells

Stratified epithelium-multiple layer of cells

Shapes: Cuboidal (cube-like) specialized secretion such as hormones

Columnar (column- or brick-like), large cytoplasmic volumes-junctions prevent
movement of molecules in between the cells. Secretion and/or absorption occurs, such
as in the small intestines)
Squamous (flat or similar to floor tiles) Leaky, such as capillaries, skin

Q: You attempted to generate small intestine cells by expressing intestinal

differentiation genes in embryonic stem cells (and knocking out “stemness” genes),
and after transplanting the cells into mice, you discovered that the cells were
squamous in shape and function instead of being columnar. Predict the
consequences of this discovery.

Stratified squamous epithelia (skin)

Loss of cells at free end

of epithelial cell layer
abrasion

Cell division of skin stem or progenitor

Basement (stem-like) cells occurs relatively
membrane close to basement membrane

4
Connective Tissue:
•  Binds and supports other tissues, many types of connective tissue cells
•  Sparse population of cells scattered throughout an extracellular matrix
• Matrix: jelly-like (adipose or fat cells) or solid (bone) web of fibers, liquid (plasma of blood)
• Matrix components generally secreted from connective tissue cells

Connective
Tissue Cell
Matrix components
Extracellular
Matrix

Distinct matrix environments, components, functions Adipose cell = fat cell

(consider adipose tissue vs bone)

Connective tissue fibers: Critical component of Tendons, Ligaments as well as many

connective tissues
Collagenous fibers: collagen, nonelastic rope-like bundle of fibrils, does not easily tear
Elastic fibers: elastin, long threads, provides a rubbery quality
Reticular fibers: collagen, thin and branched, connected to collagenous fibers, forms
a tightly woven fabric that joins connective tissue to adjacent tissue

Mesenchyme Stem Cells: normally differentiates into connective tissue cells (and nerve cells):

Patients with osteoarthritis or meniscus injuries: After surgery, approximately 50% of

the patients improve but then show deterioration within 2-5 years.

Current Clinical Trials: Surgery + Saline compared to Surgery + Stem cells

(Bone Marrow or Fat stem cells)
No results yet, but several patients showed reduced inflammation after using stem cells.

Private Stem Cell Clinics- Claim that injection of Bone Marrow stem cells directly into
joints improves the osteoarthritic condition. What are the ethical and societal issues?

5
 Nervous Tissue:
•  Senses stimuli and transmits signals throughout the organism
•  Neuron-nerve cell containing a cell body, dendrite and axon
Dendrite-transmits impulses from their tips towards the rest of the neuron
Axon-transmits impulses toward another neuron or an effector (such as a muscle)
to carry out a response.

Inputs Myelin
Dendrites Axon Sheath

Output
Effector
or target
Synapse cell
Junction between
Cell
neuron and effector
Body or between two neurons.

Neurotransmitters are released from the synapse. There are many neurotransmitters (more than
35) and examples include nitric oxide, acetylcholine, catacholamines (norepinephrine,
serotonin), or neuroactive peptides (such as endorphins).

Multiple Sclerosis (MS): Degradation of the Myelin Sheath (normally acts to

“insulate” the axon and enhance the speed of the signal down the axon)

Observed:
Mice with Inject human • Improvement of the MS-like condition
MS-like ESC into the • Loss of stem cells
defect spinal cord • Damage to Myelin Sheath mostly repaired
• Prevented more damage

Current studies-comparing the injection of human ESC to that of adult

mesenchyme stem cells

6
 Multiple Sclerosis (MS): continued

Limitations, and potential problems of stem cell therapies:

Use of human stem cells in mice

“Off Target” effects of injecting stem cells

Potential differentiation of the injected stem cells into non-nerve cells

Use of a wider range of stem cells

Stability of nerve tissue formed from stem cells

Muscle Tissue:

•  Most abundant tissue in most animals

•  Composed of long cells (muscle fibers) that can contract when stimulated by nerve impulses
•  Produce mechanical force by their contraction (highly energy consuming)
•  Cells contain many microfilaments, which is composed of actin and myosin,
arranged in parallel within the cytoplasm.

Skeletal muscle-voluntary movements of body, also called striated muscle which

is due to overlapping filaments.
Cardiac muscle-forms contractile wall of the heart, striated, branched cells,
ends of cells are joined by intercalated discs that relay signals during a heart beat
Smooth muscle-lacks striations, spindle-shaped, involuntary body activities
locations include walls of digestive tract, bladder, arteries and other organs

7
 Signal transduction pathways: Intracellular molecular cascades that allow target cells
to perceive changes in the environment and to mount an appropriate response
to extracellular stimuli

Signal transduction pathway

Response
(multiple steps in cascade)
Extracellular “Reception” of Cellular interpretation that
signal stimuli leads to an appropriate response

Examples of environmental stimuli that work through receptor mediated pathways:

Hormones/growth factors/extracellular molecules
Environmental stress such as osmotic shock and UV radiation
(work through hormone-like pathways)
Contact with other cells and the extracellular matrix

Steroid hormone action:

Steroids diffuse through the plasma membrane and into the cell
Steroids bind to an appropriate intracellular receptor (either cytoplasmic or nuclear)
Activated steroid-receptor complex is a transcription factor that binds to DNA and/or
other proteins (such as transcription factors) on the promoters of genes
which leads to an alteration in gene expression

Protein hormone action:

Protein hormone or peptide binds to a cell surface receptor

Activated hormone-receptor complex induces an interaction with other proteins
on the cytoplasmic side of the plasma membrane
Initiation of a signal transduction cascade that can involve a variety of protein interactions
with other proteins, phosphorylation events, activation of nucleotide binding proteins,
production of secondary messengers,regulated localization of proteins, etc.
Final targets of the cellular cascade include transcription factors that alter gene expression,
a variety of cellular enzymes and the cytoskeleton.