摘要
目的 :探索分离、纯化及培养成人骨髓间质干细胞 (hMSC)的最佳方法。方法 :采用Ficoll Paque淋巴细胞分离液和全血接种两种方法分离成人MSC ,筛选体外培养hMSC适合的培养基和适宜的血清含量 ,流式细胞仪检测hMSC表面抗原表达。结果 :经Ficoll Paque淋巴细胞分离液分离后 ,接种于 φ =10 0mL/L胎牛血清的L DMEM培养液 ,细胞贴壁良好 ,增殖速度快 ,hMSC在体外扩增 5代可获得 1× 10 8细胞。全血接种分离hMSC ,细胞贴壁慢、少 ,生长情况欠佳。除L DMEM ,其他类型的培养基均不适合hMSC的培养扩增。流式细胞仪检测结果显示CD2 9、CD44、CD10 5、CD166表达阳性 ,CD14、CD3 3、CD3 4、CD3 8、CD45、CD11a为阴性。结论 :用Ficoll Paque淋巴细胞分离液分离骨髓单核细胞 ,接种于 φ =10 0mL/LFCS的L DMEM培养液 。
Objective: To establish a method for isolation and cultivation of mesenchymal stem cells (MSCs) from human bone marrow. Methods: hMSCs were separated from human marrow with Ficoll Paque reagent or direct culture and expanded in different culture medium with newborn bovine serum. The proliferation and growth characteristics were observed in primary and passage culture. To detect the surface antigens, the labeled cells were analysed on a FACScan flow cytometer. Results: The isolated hMSCs comprised a single phenotypic population and displayed a fibroblast like morphology. hMSCs had a strong self renewal capacity. After P5 culture,1×10 8 cells were obtained. L DMEM was better for the proliferation and growth of MSC. These expanded attached MSCs were uniformly positive for CD29、CD44、CD105、CD166,without the expression of CD11a、CD14、CD33、CD34、CD45、CD38. Conclusion: MSCs can be satisfactorily isolated from human marrow with Ficoll Paque reagent and expanded in L DMEM culture medium.
出处
《广东药学院学报》
CAS
2002年第2期135-138,共4页
Academic Journal of Guangdong College of Pharmacy
基金
国家自然科学基金资助 (30 1 0 0 1 88)
广东省"十五"重大专项 (A 30 2 0 1 0 1 )
