人骨髓间充质干细胞不同亚群增殖分化的研究

Study on Proliferation and Differentiation of Various Subpopulations in Human Mesenchymal Stem Cells

为比较不同亚群的人骨髓间充质干细胞(human bone mesenchymal stem cell,hBMSC)自我更新和分化能力,通过获赠的人髂骨骨髓标本,联合运用密度梯度离心和差异贴壁法分离MSCs,用10μm滤膜将不同群体细胞分离,倒置相差显微镜观察不同亚群细胞的形态;流式细胞仪检测BMSC不同亚群细胞的表型;在地塞米松、Vit C、β-磷酸甘油钠作用下将不同亚型细胞向成骨细胞诱导分化,分别观察其分化能力。结果成熟的MSC,即mMSCs细胞(mature cells)呈纤维样梭形,RS细胞(rapidly MSC self-renewing cells)呈圆形。RS细胞增殖能力明显强于mMSCs。经定向诱导分化后,RS细胞向成骨细胞分化能力较mMSCs细胞强。说明RS细胞较之mMSC细胞可能是一种更原始的中胚层前体细胞,具有更强的自我更新和分化潜能。

To compare self-renewal and differentiation capacity of various subpopulations in human mesenchymal stem cells （hBMSC）, MSCs are isolated from donated human iliac marrow specimen by density gradient centrifugation and adherent screening. Cells from 80 % confluence were passed through a 10 μm filter, cell morphology is observed by inverted phase contrast microscope, phenotype and cell cycle of different subgroups were investigated by flow cytometry. Various subpopulations are induced to osteoblasts by means of Dexamethasone, Vitamin C and β- Sodium Glycerophosphate. And differentiation capacity is examined respectively. It is result mature MSC, i. e. mMSCs, are fiber-like spindle cells, while rapidly MSC self-renewing cells（ RS cells） are round. The proliferative capacity of RS cells is far better than that of mMSCs. After directional differentiation, the differentiation capacity of RS cells to osteoblasts is also better than that of mMSCs. It si concluded possibly RS cells are a kind of mesoblastic precursor cells, possessing better self- renewal and differentiation capacity.