猴骨髓间充质干细胞的分离及多向诱导分化

Isolation and induced multi-differentiation of monkey marrow mesenchymal stem cells

目的 探讨猴骨髓间充质干细胞 (MSCs)体外生长特性及多向分化潜力 .方法 以Percoll(质量密度 10 73g·L-1)非连续密度梯度离心分离出骨髓悬液中的MSCs,在非诱导条件下进行培养 ,观察细胞的形态、生长状况 .取原代体外培养3wk的MSCs分别向成软骨细胞、成骨细胞、成脂肪细胞及成肌细胞方向诱导 ,观察其诱导分化结果 .结果 原代培养的MSCs增殖缓慢 ,细胞增殖率和形态分化不均一 ,在培养过程中有多种形态的细胞出现 ,如伸展及增殖均不明显的圆形细胞 ,伸展充分但增值不明显的片状细胞、神经元样细胞 ,还可见增殖明显的克隆团状或条带状细胞群出现 .传代培养的MSCs细胞仍可见增殖情况及形态分化的多样性 ,但有“拉网”现象出现 .所培养的MSCs经诱导可向软骨细胞、成骨细胞、脂肪细胞和成肌细胞发展分化 .结论 体外培养的猴MSCs细胞具有形态分化、增殖情况多样性的特点 ,并具有多向分化潜能 .

AIM To investigate the growth features and the potential of multi differentiation of monkey MSCs in vitro . METHODS MSCs were isolated from marrow suspension by discontinuous gradient centrifugation on percoll (density: 1073 g·L -1 ) and then cultured under non inductive conditions. The morphtype and growth of the cultured cells were examined. MSCs cultured primarily 3 wk were induced to differentiate into chondroblast, osteoblast, lipoblast and myoblast respectively. And the results of the inducement were evaluated. RESULTS The growth of MSCs was slow with the heterogeneity of the multiplication rate and the morphological differentiation. There were several cell shapes during the culture. Some cells did not spread and multiply significantly, for example, the round cells. Some cells, such as lamellar cells and neurocyte like cells, were well spread, but without significant multiplication. And there were also some cells that had significant proliferation and grew into cells polarity like a clone or a band. When the MSCs were subcultured, the heterogeneity of the multiplication rate and the morphological differentiation were detected, but the 'netting' could also be found. And the MSCs could differentiate into chondroblast, osteoblast, lipoblast and myoblast respectively during the inducement. CONCLUSION The cultured MSCs of monkey had the potential of multi differentiation with the heterogeneity of multiplication and morphological differentiation.