Rhesus monkey embryonic stem cells are ideal candidates for basic research of
development biology and allogenic transplantation research because the monkey takes
advantages over rodents in close genetic and physiological relation to human.
Commonly, rhesus monkey embryonic stem cells are maintained on feeder cells, but
feeder cells limit the research and application of rhesus monkey embryonic stem cells.
So, it is necessary to establish a feeder-free culture system of rhesus monkey
embryonic stem cells. Here we demonstrate a successful feeder-free culture system for
IVF3.2, which is a rhesus monkey stem cell line derived from in vitro fertilized
embryo and established by our lab. In this system, IVF3.2 is cultured on Matrigel in
medium conditioned by MEF and subcultured by trypsinization. We mainly studied
the capability of cell growth and differentiation in the feeder-free culture system, and
investigated the cell cycle distribution and the cell viability after cryopreservation in
two culture system. The results are shown as follows:
1) In the feeder-free condition, IVF3.2 maintained the two basic
characteristics of embryonic stem cells including the properties of
self-renewal and pluripotency.
2) There is no significant differentiation of the cell cycle distribution
between the cells cultured on feeders or off feeders. More than half of the
growing rhesus monkey embryonic stem cells are in S phase in both
culture conditions.3) IVF3.2 cultured off feeders in this system can effectively survive freezing
and thawing and could be cultured for prolonged periods. The flow
cytometry analysis revealed that the percentage of viable cells after
standard slow freezing is up to 83% for the cells cultured off feeders,
which is significantly higher than the cells cultured on feeders.
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