胎盘间充质干细胞脐血血浆培养体系的建立

Establishment of cord blood plasma culture system for placenta-derived mesenchymal stem cell expansion

目的建立并优化适用于临床级胎盘源间充质干细胞的脐血血浆体外培养体系。方法采用酶消化法分离胎盘间充质干细胞,分为胎牛血清组(10%胎牛血清+DMEM/F12)、脐血血浆组(10%脐血血浆+DMEM/F12+1 000 IU/L肝素)和无血清组(Stem ProMSC SFM CTSTM);经培养扩增后,采用流式细胞术检测间充质干细胞表面标志,并诱导细胞向成脂肪、成骨分化,比较3组培养扩增所得间充质干细胞的差异。结果 3组细胞的增殖速度并无明显差异;脐血血浆组、胎牛血清组及无血清组培养扩增所得的细胞均表达CD90、CD29、CD44、CD49e、CD105、CD73,不表达CD34、HLA-DR、CD45;而且均具有向成脂、成骨分化的能力。结论脐血血浆可取代胎牛血清或无血清培养基用于胎盘间充质干细胞的培养扩增,以此获取可供临床应用的胎盘间充质干细胞。

Objective To establish in vitro culture system of cord blood plasma optimized for clinically functional placenta- derived mesenchymal stem cells. Methods Collagenase digestion was applied for isolation of mesenchymal stem ceils （MSCs） from placenta. MSCs were divided into 3 separate groups based on different culture media, namely fetal bovine serum, cord blood plasma and serum - free medium. After cultured expansion, MSC surface markers were de- tected by flow cytometry. And MSCs were induced to differentiate into adipocyte or osteoblast. Differences of MSCs among the 3 groups were compared. Results There was no difference among the three groups in proliferation rate. CD90, CD29, CD44, CD49e, CD105 and CD73 were positive in all of the three groups while CD34, HLA - DR, CIM5 were negative. Cells in all of the three groups were able to differentiate into adipocyte and osteoblast. Conclusion Both fetal bovine ser- um and serum - free medium can be replaced by cord blood plasma in the in vitro cultured expansion of placenta - derived MSCs for clinical purposes.