核心结合因子α1过表达促进脂肪组织来源干细胞向成骨细胞分化

目的 检测核心结合因子α1(Cbfα1)的特异性成骨作用以及脂肪组织来源干细胞的成骨分化潜能.方法 采用Cbfα1重组的腺病毒载体感染脂肪来源干细胞,通过间接免疫荧光的方法检测脂肪来源干细胞中Cbfα1的表达.Cbfα1转染脂肪组织来源干细胞后1、3、7 d利用RT-PCR检测成骨特异性基因和成脂特异性基因的表达变化.同时通过检测Cbfα1转染后7、10 d脂肪来源干细胞碱性磷酸酶活性的变化,观察Cbfα1过表达后诱导脂肪组织来源干细胞向成骨细胞分化.结果 转染后脂肪组织来源干细胞细胞数目增长一倍.在特定诱导条件下,可向成脂细胞、成骨细胞分化.Cbfα1重组的腺病毒载体对脂肪组织来源干细胞的转染效率为82.4%.转染后第1、3、7天脂肪组织来源干细胞中骨钙素、骨桥素、Ⅰ型胶原的表达增强;脂蛋白脂酶的表达逐渐下降.转染Cbfα1后,脂肪组织来源干细胞中碱性磷酸酶活性显著增加.结论 核心结合因子过表达促进脂肪组织来源干细胞向成骨细胞分化、抑制成脂分化.

Overexpression of core binding factor α1 enhancing osteoblastic differentiation in adipose-derived stem cells

Objective To determine whether mouse core binding factor α1(Cbfα1) overexpression enhancing osteoblastic differentiation in adipose-derived stem cells(ADSCs). Methods ADSCs were harvested from SD rats and transduced with the recombinant adenovirus carrying Cbfα1 gene. Untransduced cells and cells transduced with adenovirus carrying the enhanced green fluorescence protein(Ad-EGFP)gene served as controls. Cbfα1 expression was assessed by immunofluorescence. Alkaline phosphatase (ALP)activity was assayed in 7 and 10 d. Results Overexpression of Runx2 inhibited adipogenesis,as demonstrated by suppression of LPL expression. Moreover,ADSCs transduced with Ad-Runx2 underwent rapid and marked osteoblast differentiation as determined by osteoblastic gene expression,alkaline phosphatase activity. Conclusions Cbfα1 overexpression enhances osteoblastic differentiation in adiposederived stem cells.