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| A Study on Six Kinds of Cell Culture System for Mouse Embryonic Stem Cells |
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| DOI： |
| KeyWord:Mouse Embryonic stem cells Culture system |
| Huang-Bing,Huang Wen-ge,Zhong Nu-qi,Chen Ying-qing,Chen Xi-gu |
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| Abstract: |
| Objective To establish six kinds of stable embryonic stem cell
culture system. Methods Mouse embryonic stem cells (ES D3 line) are maintained in following culture
systems The primary mouse embryonal fibroblasts (MEF) in serum medium and MEF in
serum-free medium as well as SNL (mouse fibrob last subline ) in serum medium all together
consist three kinds of feeder layer dependent culture systems. The serum medium and
serum-free medium supplemented w ith recombinant murine leukemia inhibitory factor (mLIF)
respectively as well as Buffalo Rat Liver (BRL) conditioned medium consist three varied
feeder-free lay er culture systems. The ES D3 cells are passaged 10 times in vitro and
their clo ne are observed morphologically. In order to identify ES cell pluripotent and un
differentiation on the above mentioned systems, we tested the alkaline phosphata se (AKP)
staining which is a marker of the undifferentiated cells and the experi ment of ES D3
cells differentiation in nude mice were investigated. Results On both feeder layer systems and feeder free systems, the
typical ES cells clone shape r evealed as follows nested aggregation (clone) with clear
edge and smooth surfac e as well as close arrangement within the clone. Strong positive
AKP reactive c ells are observed. On the other hand. the teratomas made up with various
tissue s are formed in nude mice. Conclusion Each of the six culture systems can be us ed to promote
the growth of ES D3 and maintain their undifferentiation and pluri potentiaty, which lay a
solid foundation for the application of ES cells. |
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