Objective To compare proliferative activity, osteogenic and adipogenic differentiation capacity among different passage numbers of human amniotic mesenchymal stem cells( hAMSCs ). Methods The hAMSCs were isolated from human placental basal decidua hy collagenase and trypsinase digestion methods. The superficial molecular markers of these cells were detected hy flow cytometry. After serial passage , passage 3 ,5 ,7 ,9( P3 , P5 , P7 , P9 )were singled out for the assay of proliferative activity by MTT method and the findings were delineated as growth curve. Furthermore, these cells of above four passages were respectively induced by adding osteogenic and adipogenic inducers. 3 w later, intracellular lipid deposition was verified by oil red O staining and extracellular calciumphosphate precipitates were stained with alizarin red. The positive rate of osteogenic staining and adipogenic rate were compared among different passage numbers of hAMSCs. Results hAMSCs of P9 still showed a colony - like growth property , but their proliferative activity was down regulated. Intracellular lipid droplets decreased from P5( P ＜ 0.01 ). and extracellular calciumphosphate did from P7( P＜ 0.01 ). Conclusion hAMSCs can keep growth characteristics of mesenchymal stem cells during serial passage. However,multipotential differentiation capacity is varied among different passage numbers.%目的 比较不同传代倍数的人羊膜来源的间充质干细胞(hAMSCs)的增殖活性以及成骨、成脂的分化能力.方法 采用胶原酶和胰蛋白酶法分离人胎盘羊膜来源的MSCs,测定分离细胞的表面分子标志.对第3、5、7、9代hAMSCs绘制细胞生长曲线,分别加入成骨和成脂诱导剂,诱导3 w后行茜素红和油红O染色,比较不同传代倍数的hAMSCs成骨染色阳性率和成脂率.结果 hAMSCs传代至第9代时仍呈克隆样生长,但细胞增殖活力有下降;在第5代后成骨分化能力下降(P＜0.01),而传代至第7代后,成脂分化能力才有下降(P＜0.01).结论 hAMSCs在连续传代中能保持MSCs的生长特性,但多向诱导分化能力在不同的传代倍数中有一定差别.
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