ABSTRACT
This test aim at exploring and optimizing the isolation and culture methods of chicken amniotic fluid stem cells (CAFSCs) in vitro, characterizing CAFSCs and inducing CAFSCs to differentiate into three layers. Methods and results include that more CAFSCs are gained via thermoelectric method with lower cost. Groping and optimizing culture system abiding by the idea of simulating the chicken body environment, involving adding proportion of chicken serum and some growth factors. CAFSCs can be transferred to 34 passages nowadays. Surface marker Oct4, CD105, nanog, CD73 and SSEA-4 on CAFSCs are detected positive with immunofluorescence histochemistry, Gene CD44, CD29, CD73 and SH2 are detected positive with RT-PCR. We carry karyotype analysis out and find that chromosome is 2n=78 and shows normal. CAFSCs in mid period of subculture propagate fastest from cell cycle examination on Flow Cytometer and growth curve using CCK-8 kit on ELIASA. CAFSCs are induced to neuron and epithelium (epiblast), adipocyte (mesoblast), hepatocyte (entoblast).
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