腺病毒表达载体对骨髓间充质干细胞分化能力的影响  

Effect of adenoviral vectors on the differentiation potential of human bone marrow mesenchymal stem cells

作　　者：单志新[1] 林秋雄[1] 李晓红[1] 邓春玉[1] 周志凌[1] 黄薇[1] 黄晓忠[1] 余细勇[1] 

机构地区：[1]广东省人民医院医学研究中心/广东省心血管病研究所,广东广州510080

出　　处：《南方医科大学学报》2008年第7期1132-1135,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(30300421;30672077);广东省自然科学基金(06020831)~~

摘　　要：目的研究腺病毒感染骨髓间充质干细胞(MSC)对MSC分化潜能的影响。方法利用表达绿色荧光蛋白(GFP)的重组腺病毒感染传至3代的人MSC,分别在感染后第2、4、8和16天收集细胞,提取总RNA,利用RT-PCR检测感染前后MSC中内胚层标志基因CYP51、中胚层标志基因SM22α、外胚层标志基因槽蛋白(nestin)、多分化潜能标志基因OCT4和可变剪切因子基因nPTB的表达。在腺病毒感染MSC7d后,利用成脂肪诱导剂继续诱导培养14d,用油红O染色观察MSC向脂肪细胞分化情况。结果RT-PCR检测显示,MSC中CYP51、SM22α、nestin、OCT4和nPTB在腺病毒感染后2、4、8和16d时仍有表达。腺病毒感染7d后的MSC仍可向脂肪细胞分化,且分化效率同正常MSC一致。结论腺病毒载体感染MSC后,不会影响MSC的分化能力,可以作为MSC诱导分化机制研究的基因表达载体。Objective To investigate the effect of adenoviral vector infection on the differentiation potential of human bone marrow mesenchymal stem cells （hMSCs）. Methods The third-passage hMSCs were infected with the recombinant adenovims expressing green fluorescence protein （GFP） for 2, 4, 8 and 16 days. RT-PCR was used to detect the mRNA expression of endodermal marker CYP 51, mesodermal marker SM22α, ectodermal marker nestin, pluripotent marker oct-4 and the alternative splicing factor nPTB. Seven days after adenovirus infection, the hMSCs were cultured in the presence of adipogenic agents for 14 days, and the adipose cells differentiated fi＇om hMSCs were detected with oil red O staining. Results Compared with normal hMSCs, the cell infected with the adenovirus for 2, 4, 8 and 16 days showed no obvious down-regulation of CYP51, SM22α, nestin, OCT4 or nPTB. The hMSCs 7 days after adenovirus infection were induced to differentiate into adipose cells, with a similar differentiation rate to that of normal hMSCs. Conclusion The differentiation potential ofhMSCs is not affected by adenovirus infection, suggesting that adenovirus can be used as the gene delivery vector in MSC differentiation studies.

关 键 词：骨髓间充质干细胞 腺病毒 载体 分化 

分 类 号：Q78[生物学—分子生物学] Q254