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180 EFFECT OF RAT CARTILAGE ESSENCE ON OMENTUM CELLS CULTURED BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS
Zahra Goudarzi et al.
NMRI ratis extracted and summarized. Some lab stud-
ies have shown that if mature chondrocytes are cultured
in simple dishes, lose their differentiation rapidly and
their physiological activities decrease (Ng 2005). One
of the important conditions for creating culturing is
using micromass (Johnstone,1998). Weused this method
for creating suitable cellular mass, which is increasing
number of cells in level unit.
Thereafter, cells phenotype and genotype was con-
rmed with various evaluations such as ow cytometry.
Accordingly, in human and other rodents such as rat, sur-
face antigens related to these cells were investigated by
ow cytometry and using monoclonal antibodies. What
today is important in identi cation of MSCs, is if they
have broblast morphology and are negative in terms
of hematopoietic stem cells, they can be presented as
MSCs (Barry, 2004). Expressing speci c markers of MSCs
includes CD90 and CD44 and lack of expressing special
marker of hematopoietic cells is con rmed as CD45 in
separated cells from the omentum, which is aligned with
Pancholi et.al results (Pancholi et al 2010 Li, 2005).
In this research cartilage essence was used for cre-
ating differentiation induction instead of using com-
mon growth factors, since using growth factors such as
TGF can have unknown effects on other cells and have
tumorigenic effects. However, using natural essences in
addition tolacking the above mentioned shortcomings
can have the advantage of lesser transplant rejection due
to tissue similarities and cells differentiate with more
similar conditions with their natural den. Treatment
with cellular essence, is a new strategy for cells differ-
entiation which is able to transform a type of somatic
cell into another. MSCs affected by cartilage essence
induction can nd chondrocyte cells morphology. For
instance cells took an oval shape which is a characteris-
tic of chondrocyte cells (Shamblott, 1998). Then matrix
construction begins to separate chondroblasts. Differen-
tiation occurs from the center toward exterior. There-
fore, more central cells have chondrocytes characteris-
tics, however, ambient cells typically are chondroblasts
(Liu, 2010).
Various dilutions of cartilage essence were used, after
mesenchymal cells were treated by cartilage essence
with 50 landar dose, signi cant changesoccurred from
the fourth day onward. In some regions cellular colonies
were created. With an increase in number of cells, they
absorb cellular extras. And cells move toward oval and
round formation, which means they are chondroblasts
which later use of speci c Alison blue and Toludine blue
coloring con rmed this notion. These cells seem com-
pletely alive and active and replication process still con-
tinues in them. These change occur while in other groups
under study, their cells still remain between chondro-
cytes morphology phase and spindly and lengthy mor-
phology which is speci c tp broblasts and with passage
of time face senescence and apoptosis.
Collagen type 2 is the most abundant existing col-
lagen in cartilage. Collagen type 2 contains speci c
sequences to connect to the cell and as a result has
a high interaction with the cell. It can be suitable for
maintaining cartilage cells phenotype and activities (Li,
2005, Zambrano, et al, 1982). In this experience we were
able to con rm production of collagen type 2 through
immunohistochemistry in cartilage cells derived from
omentum MSCs of NMRI breed rat after 21 days from
the start of differentiation induction. Therefore, consid-
ering cellular mass and morphological changes in the
cultured stem cells it seems that we succeeded in pro-
ducing chondroblasts.
CONCLUSION
This research is carried out to investigate inducing
effects of cartilage essence obtained from rat on cul-
turing omentum cells through micromass with reducing
serum amount and understanding and interpreting the
results. Results are the indication of the cartilage essence
effect on omentum`s MSCs in the absence of growth fac-
tors, common induction and their differentiation with
chondrocytes. It seems that our method of separating
omentum`s mesenchymal cells and producing cellular
lnie was completely successful and powerful and have
appropriate and acceptable progress toward differentia-
tion, specially creating a compression and production of
chondroblasts. Considering the results that Dubler et.al
have obtained from culturing on collagen and results
that Tuli and Okafor obtained regarding culturing with
tissue engineering using collagen type 2, we can con-
clude that the cartilage essence used in this research as
an induction factor can intensify differentiation process
in cells due to existence if collagen in itself with the help
of creating and maintain cellular connections.
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