Establishment Of Chicken Amniotic Fluid Stem Cells Line And Effects Of Ginsenoside To Its Proliferation

This test aimed at exploration the isolation,culturing methods of Chicken Amniotic Fluid Stem Cells(CAFSCs) and filtrating,optimizing its culturing system in vitro,Then the effects of ginsenoside to CAFSCs proliferation were researched.The fertilized eggs between seven and nine days were selected, supercentrifugation method and thermoelectric method were used to isolate CAFSCs;We used whole culture medium that contained a certain proportion of chicken serum and growth factor;immunofluorescence histochemistry,RT-PCR were applied to identify surface molecular mark of CAFSCs;ginsenoside on the different level of concentration was used to handle CAFSCs separately,which would be tested to get its proliferation messages with CCK-8,EdU-EU and RT-PCR.More CAFSCs were gained via supercentrifugation method and thermoelectric method with lower cost;CAFSCs presented strip-type,adhered and Vigorous in virto,which could be transfered to thirty-four passages nowadays;Surface marker Oct4,CD105,nanog,CD73 and SSEA-4 on CAFSCs were detected positive with immunofluorescence histochemistry;Surface marker CD44,CD29,CD73 and SH2 were detected positive with RT-PCR;When observeing under inverted microscope,we could find cells spreading all around behind the ginsenoside handle;We detected the proliferation of passage five CAFSCs with CCK-8,drew growth curve and carrued analysis with SPSS,then found that OD value is higher along with concentration increasing continuously(p<00.01) in a specific concentration range.We detected the proliferation of passage five CAFSCs with EdU-EU by image analysis,the higher concentration,the more cell quantities of proliferation in a specific concentration range;We detected the proliferation of passage twenty five CAFSCs with RT-PCR,then carried gray intensity analysis,the higher concentration,the more expression of proliferation gene NIFK,EGFR,C-myc in a specific concentration range.It has been reported that this experentment is the first case to isolate and culture bird species amniotic fluid stem cells successfully nowadays.Experments indicate that CAFSCs can beobtained easily,cost lower, own stronger proliferation capacity in vitro and suitable for isolation and culture on a large scale in vitro.It offers a better choose for stem cells.Ginsenoside can promote CAFSCs proliferation,which will advance its application in medicine.