永生化人骨髓基质细胞的表面抗原及多向分化研究

Biological characteristics of human bone marrow stroma cells immortalized by human telomerase reverse transcriptase gene

目的研究本所转入人端粒酶逆转录酶基因的人骨髓基质细胞(hMSC-TERT细胞)的生物学特性。方法复苏hMSC-TERT细胞第30代,扩增,流式细胞仪检测细胞表面抗原;进行体外脂肪细胞、成骨细胞定向诱导,并分别进行油红O染色、碱性磷酸酶染色及von Kossa染色。结果hMSC-TERT细胞表面抗原检测CD34、CD45、CD44、CD106、CD166均为阴性;脂肪细胞定向诱导后油红O染色见胞浆内有红色脂肪颗粒,成骨细胞定向诱导后见胞浆内有碱性磷酸酶染色阳性黑褐色颗粒,von Kossa染色见有钙结节形成。结论hMSC-TERT细胞依然保留骨髓基质干细胞的自我更新特性及多向分化潜能。

Objective To investigate the biological characteristics of human bone marrow stroma cells immortalized by human telomerase reverses （bMSC-TERT）. Methods The 30^th passage hMSCs-TERT were amplified after recovery of cryopreservation. The surface antigenic features of the cells were analyzed by flow cytometry. Differentiation induction of adipose cells and osteoblasts was performed in vitro. The induced hMSCs-TERT were examined by Oil red-O staining, alkaline phosphates staining and yon Kossa staining. Results The hMSC-TERT was negative for CD34, CD45, CD44, CD106 and CD166. After the induction, accumulated cytoplasmatic red lipid droplets were clearly visible in the adipose cells by Oil red O staining, and alkaline phosphates-positive pitchy grains formed in the intracytoplasm and calcium nodes were found by yon Kossa staining in the osteoblasts. Conclusion The hMSC-TERT still retains the self-renewing characteristics of bone marrow stromal stem cells and pluripotent differentiation potential.