不同来源人羊膜间充质干细胞的混合培养

Mixed culture of human amnion-derived mesenchymal stem cells from different sources

背景:人羊膜间充质干细胞可诱导分化为神经样细胞,但在培养过程中发现干细胞增殖的数量不足。目的:为了获得足够数量的人羊膜间充质干细胞,观察人羊膜间充质干细胞混合培养的生长情况及经碱性成纤维细胞生长因子诱导人羊膜间充质干细胞向神经样细胞的分化情况。方法:将两个不同胎盘来源的羊膜分离培养、混合,采用细胞分离和培养技术获取人羊膜间充质干细胞,分离人羊膜间充质干细胞后传代培养,加入碱性成纤维细胞生长因子诱导分化。当细胞传至第3代时,随机取培养的6份人羊膜间充质干细胞(半量)为A组,6份人羊膜间充质干细胞(半量)为B组,将A、B组各剩余的半量人羊膜间充质干细胞混合为C组。3组细胞浓度均为1.0×107L-1。以活细胞计数和MTT比色法比较3组细胞扩增数量,以免疫组织化学法检测羊膜间充质干细胞神经胶质细胞标志物、神经元特异性烯醇化酶和巢蛋白的表达。结果与结论:混合人羊膜间充质干细胞之间有互相促增殖的作用。人羊膜间充质干细胞具有较强的可朔性,经碱性成纤维细胞生长因子诱导的人羊膜间充质干细胞可表达神经胶质细胞标志物、神经元特异性烯醇化酶和巢蛋白。

BACKGROUND：Human amnion-derived mesenchymal stem cells（HAMs） can be induced to differentiate into neuron-like cells,but the number of stem cells in the training process is insufficient.OBJECTIVE：Observed HAMs the growth of mixed culture,and induced HAMs to the nerve-like cells by basic fibroblast growth factor（bFGF） to obtain a sufficient number of HAMs.METHODS：Amnions from two placentas were separated and cultured to obtain HAMs,which were induced differentiation by adding bFGF.When cells were at the 3rd passage,cultured 6 HAMs（half dose） were randomly taken for the A group,6 HAMs（half dose） were taken for the B group,the remaining half of the amount of the A,B group HAMs mixed as C group.Cell densities of 3 groups were 1.0×107/L.The cells proliferations of 3 groups were compared by live cell count and MTT colorimetric.The expressions of GFAP,neuronspecific enolase（NSE） and nestin were detected by immunohistochemistry.RESULTS AND CONCLUSION：Mixing HAMs have mutual effect on the proliferation.HAMs have a strong retrospective,which can express GFAP,NSE,and nestin induced by bFGF.