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目的对人羊水标本进行体外分离培养,建立人羊水来源干细胞的体外培养体系,对其生物学形状进行研究。方法贴壁法体外分离获得人羊水来源干细胞,多次传代扩增后,采用流式细胞仪和RT-PCR技术检测细胞表面抗原的表达。结果羊水干细胞原代生长较慢,传代后生长迅速,体外倍增时间约36h,流式细胞仪检测证实细胞表达CD29、CD44、CD105等间充质干细胞标志,不表达造血干细胞标志CD45和CD133。RT-PCR检测显示羊水干细胞表达Oct-4、Nanog基因。结论实验成功分离获得羊水中具有干细胞性质的细胞群,采用贴壁法分离获得的干细胞体外增殖能力强,表达间充质干细胞表面标志,符合间充质干细胞的特点。
Objective To isolate and culture human amniotic fluid samples in vitro and establish a culture system of human amniotic fluid derived stem cells in vitro and to study their biological shapes. Methods Human amniotic fluid derived stem cells were isolated and isolated in vitro. After multiple passages, the cell surface antigens were detected by flow cytometry and RT-PCR. Results The primary growth of amniotic fluid stem cells was slow, and grew rapidly after passage. The doubling time was about 36 h in vitro. Flow cytometry confirmed that the cells expressed markers of mesenchymal stem cells such as CD29, CD44 and CD105, but not hematopoietic stem cells markers CD45 and CD133. RT-PCR showed that amnion stem cells expressed Oct-4 and Nanog genes. CONCLUSION: The stem cells isolated from amniotic fluid were successfully isolated from the amniotic fluid. The stem cells obtained by adherence method were capable of proliferating in vitro and expressing the surface markers of mesenchymal stem cells, which accorded with the characteristics of mesenchymal stem cells.