重组hCDMP1腺病毒转染骨髓间充质干细胞对其向软骨分化的影响

目的探讨人软骨源性形态发生蛋白1(CDMP1)基因转染对骨髓间充质干细胞(BMSCs)增殖及分化的影响。方法采用腺病毒转染方法将重组人CDMP1(hCDMP1)基因转入体外培养的兔BMSCs,用免疫印迹法(Western blot)检测hCDMP1蛋白质的表达,并通过检测细胞增殖能力(MTT法)、Ⅱ型胶原(ColⅡ)以及蛋白多糖的表达,分析转染hCDMP1对BMsCs增殖、分化的影响。结果 hCDMP1蛋白在基因转染细胞内得到正确表达;hCDMP1基因转染组和对照组相比,ColⅡ、蛋白多糖表达水平显著增高(P<0.05),而细胞增殖能力无明显变化(P>0.05)。结论外源基因转染可以使BMSCs表达有生物活性的hC-DMP1,高表达的hCDMP1可以促进BMSCs向软骨表型分化,但对细胞增殖无明显影响。

Effects of adeno-hCDMP1 transfection on in vitro chondrogenic differentiation of rabbit bone marrow mesenchymal stem cells

Objective To investigate the effects of gene transfaction with human cartilage-derived morphogenetic protein 1(CDMP1) on the differentiation and proliferation of rabbit bone marrow mesenchymal stem cells(BMSCs).Methods BMSCs were obtained from New zealand rabbits.Exogenous recombinant hCDMP1 was transfected into BMSCs with adenovirus method.Then hCDMP1 expression at protein level was measured by western blot method.Expression of Type 1I collagen(ColⅡ),proteoglycan and growth of the cells were all measured by biological methods to evaluate the effects of gene transfer on the differentiation of rabbit BMSCs.Results After hCDMP1 gene transfection,BMSCs expressed hCDMP1 protein,and compared with the control groups,expression of proteoglycan and ColⅡincreased significantly,but no significant difference appeared in cell proliferation.Conclusion Gene transfer with hCDMP1 is an effective way to enhance the expression of CDMP1 at protein.The expression of heterogenetic hCDMP1 gene can induce BMSCs′differentiation to chondrogenic cells.But the gene transfection has no obvious effects on the proliferation of BMSCs.