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《Chinese Journal of Reparative and Reconstructive Surgery》 2015-01
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ZHANG Ziji;KANG Yan;ZHANG Zhiqi;YANG Zibo;FANG Shuying;SHENG Puyi;HE Aishan;FU Ming;LIAO Weiming;Department of Joint Surgery,First Affiliated Hospital of Sun Yat-sen University;  
Objective To investigate the micro RNA(mi RNA) expression profile during chondrogenic differentiation of human adipose-derived stem cells(h ADSCs),and assess the roles of involved mi RNAs during chondrogenesis. Methods h ADSCs were harvested and cultured from donors who underwent elective liposuction or other abdominal surgery. When the cells were passaged to P3,chondrogenic induction medium was used for chondrogenic differentiation. The morphology of the cells was observed by inverted phase contrast microscopy. Alcian blue staining was carried out at 21 days after induction to access the chondrogenic status. The expressions of chondrogenic proteins were detected by ELISA at 0,7,14,and 21 days. The mi RNA expression profiles at pre- and post-chondrogenic induction were obtained by microarray assay,and differentially expressed mi RNAs were verified by real-time quantitative PCR(q RT-PCR). The targets of the mi RNAs were predicted by online software programs. Results h ADSCs were cultured successfully and induced with chondrogenic medium. At 21 days after chondrogenic induction,the cells were stained positively for alcian blue staining. At 7,14,and 21 days after chondrogenic induction,the levels of collogen type II,Col2a1,aggrecan,Col10a1,and chondroitin sulfate in induced h ADSCs were significantly higher than those in noninduced h ADSCs(P0.05). Eleven differentially expressed mi RNAs were found,including seven up-regulated and four down-regulated. Predicted target genes of the differentially expressed mi RNAs were based on the overlap from three public prediction algorithms,with the known functions of regulating chondrogenic differentiation of stem cells,selfrenewal,signal transduction,intracellular signaling cascade,and cell cycle control. Conclusion A group of mi RNAs and their target genes are identified,which may play important roles in regulating chondrogenic differentiation of h ADSCs. These results will facilitate the initial understanding of the molecular mechanism of chondrogenic differentiation in h ADSCs and subsequently control h ADSCs differentiation,and provide high performance seed cells for cartilage tissue engineering.
【Fund】: 国家自然科学基金资助项目(81171709;81301558;81371941;81201388);; 教育部高等学校博士点基金新教师类资助项目(20130171120074);; 广东省自然科学基金博士启动项目(S2013040016269)~~
【CateGory Index】: R329.2
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