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Peter Ma

Tayyub Ali
Muhammad Fahad Khan

Kacper Rusiak
Jaleed Ahmed

ISOLATION AND CULTURE OF HUMAN


OSTEOBLASTS

YOUTUBE VIDEO
http://www.youtube.com/watch?v=78RB

pWSOl08#t=58
0.58s - 1.56s

IMPORTANCE OF OSTEOBLASTS

Osteoblasts are mononuclear cells that are vital to


human bone health

They assist the body in the areas of bone function,


repair and growth

Bone is a dynamic tissue that continually undergoes a

process called bone remodeling consists of the tissue


being broken down and then built back up

OSTEOBLASTS FUNCTION

Osteoblasts: (Greek: osteo bone and blast germ)

Osteoblasts main function is to synthesize and mineralize bone


allowing it to support the human body

Osteoblast cells have primarily been used in investigating the


skeletal systems:

the effects of inflammatory mediators, regulation of bone metabolism, and


the study of chemicals associated with physical trauma

APPLICATIONS

Osteoblasts have also been used in conjunction with carbon

nanotubes

Carbon nanotubes possess extraordinary properties in increasing


the strength of hydroxylapatite (HA)

HA is used in medical appliances placed within a patients body


to replace amputated bones or as a coating on prosthetic limbs

APPLICATIONS

Recent research has found how HA reinforced with carbon


nanotubes would behave within the body through the use of
osteoblasts cells

Figure 1: Flexible HA approximately having mineral


to organic matrix ratio of human bone
Source:http://en.wikipedia.org/wiki/Hydroxylapatite

DISTRIBUTORS

There are two major distributors for osteoblasts cells;


Cell Applications Inc and PromoCell

The average price for one ampoule of more than 500,000

cells can go for approximately 700-900$

BONE CELLS

There are four different types of bone cells that


continuously maintain bones.
These are: osteoclasts, osteoblasts, osteocytes, and
lining cells.

Figure 2: Bone cells


Source:Eijken,M. (2007). Human Osteoblast Differentiation and Bone Formation: Growth Factors,
Hormones, and Regulatory Networks

MESENCHYMAL STEM CELLS

Human osteoblast cells are a highly specialized type of


cell derived from mesenchymal stem cells (MSCs)

MSCs cells are multipotent cells that can differentiate

into a variety of cell types such as fibroblasts,


myoblasts

Mainly found in the bone marrow

MSCS CELLS POTENCY

Figure 3: MSCs cell differentiation


Source: http://www.discoverymedicine.com

HOW OSTEOBLASTS FORM

There needs to be a certain signal for MSCs cells to


generate osteoblasts. The two important transcription
factors that play an important role in MSCs cells being
converted to osteoblasts are Runx and Osterix

Through tests done on mice, it was discovered if they had


a gene deficiency of Runx2 and Osx then they had defective
bone formation and lack of mature osteoblasts.

Once the MSCs cells are committed to become osteoblasts,

they are called osteoprogenitor cells

Mesenchymal
Cells

WNT
Signalling

Skeletal
Precursor

RUNX2

BMP & WNT


Injection

Osteoprogenitor

Preostioblast

RUNX2 &
Osterix
Transcription
Factors
Initiate OBG

Osteoblasts

DETECTION OF ALKALINE PHOSPHATE IN PROLIFERATING


OSTEOBLASTS
Prepare
Solutions &
Buffers

Wash the
Cells

Fixation of
the Cells

Stain the
Cells

Wash the
Cells

OSTEOBLAST MINERALIZATION
Seed
Osteoblast

Grow
Osteoblast

Induce
Osteoblast

Differentiation
Culture of
Induced
Osteoblasts

FERMENTATION OF OSTEOBLASTS
Osteogenic differentiation

-> Seed Mesenchymal stem cells


-> Grow the cells
-> Induce mesenchymal cells with differentiation medium
-> Incubate for 21 days
-> Change medium every 3 days to populate cell culture

OSTEOBLAST DETECTION
1. Prepare solution
2. Washing Cells
3. Fixation of Cells
4. Stain the cell
5. Wash and analyze

NON-FERMENTATION
-> Use osteoblasts to derive a new product
-> Mineralization of osteoblasts
-> Osteoblasts synthesize collagen, produce calcium and
phosphate
-> hydroxyapatite mineral (Ca10[PO4]6[OH]2
-> Osteoblasts are used to create bone matrix
-> Mineral content in bone is hard to pinpoint

CANADIAN REGULATIONS
-Osteoblasts research done mostly on rat and dog osteoblast cells.
-Potential tumorigenic hazards associated with heavy metal
internalization in humans
-Testing of the bioengineered bones and cartilages as well as
osteoblasts injections still not allowed in humans.

-The use of human osteoblasts descended from mesenchymal stem


cells is much less controversial than the use of embryonic stem
cells
-The immunology issues (Physical activity, Age, Disease
environment)

The End