骨髓基质细胞亚群中多能性相关因子的表达和作用

The expression of pluripotency markers in the subpopulations of human bone marrow stromal cells

目的通过检测体外培养人骨髓基质细胞(BMSCs)各细胞亚群中多能性相关因子Sox2、c-Myc和hTert表达水平的影响,探讨BMSCs各细胞亚群多能性和重编程能力的差异。方法采用有限稀释法克隆化培养BMSCs,从快速生长克隆、慢速生长克隆和混合培养BMSCs中各选择3个样本,免疫荧光染色检测Sox2、c-Myc和hTert表达,实时荧光定量聚合酶链反应检测Sox2、c-Myc和hTert mRNA表达,进行统计学分析。结果 Sox2在快速生长BMSCs细胞克隆为细胞核强表达,在其他各组为胞浆表达,Sox2 mRNA在快速生长细胞克隆表达显著高于其他各组(P<0.05)。c-Myc和hTert呈现相似的表达模式,在快速生长细胞克隆和混合培养BMSCs均为细胞核强表达,在慢速生长细胞克隆为胞浆表达,而三组间c-Myc和hTert mRNA表达水平均差异无统计学意义(P>0.05)。结论快速生长BMSCs细胞克隆中多能性相关因子Sox2表达高于慢速生长细胞克隆,提示细胞亚群的克隆形成能力可能与细胞多能性和重编程能力正相关,Sox2可能是细胞未分化性能的关键调控因子。c-Myc和hTert可能存在协同作用。

Objective The purpose of this study was to investigate the expression of pluripotency markers Sox2, c-Myc and hTert in the subpopulations of human bone marrow stromal cells with in vitro culture. Methods Human BMSCs were culture with limiting dilution method, 3 samples were selected from fast-, slow-growing clones and mixed culture BMSCs respectively. The expression of Sox2, c-Myc and hTert was investigated by immunofluorescent staining and quantitative Realtime PCR analysis. Statistics analysis was applied. Results Sox2 showed positive nucleus expression in fast- growing clones, and cytoplasm expression in other groups. Sox2 mRNA displayed significantly higher expression level compared with other groups （P〈 0.05）. c-Myc and hTert showed similar expression pattern which demonstrated a nucleus location in fast-growing clones and mixed cultured BMSCs, albeit cytoplasm staining in slow-growing clones. Whereas the c-Myc and hTert mRNA showed no difference among three groups （P 〉 0.05）. Conclusions BMSCs from fast-growing clones demonstrated higher expression of reprogramming markers compared with slow-growing clones, indicating that colony forming efficiency may be related with the pluripotency and reprogramming capability of BMSCs. c-Myc and hTert may interact cooperatively during this process.