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当前位置: 首页 > 文章 > 牛诱导性多能干细胞向心肌细胞的诱导分化 西北农林科技大学学报(自然科学版) 2010 (10) 1-6
Position: Home > Articles > Induced differentiation of cardiomyocytes from bovine induced pluripotent stem cells Journal of Northwest A & F University(Natural Science Edition) 2010 (10) 1-6

牛诱导性多能干细胞向心肌细胞的诱导分化

作  者：
陈冬梅;吕长荣;辛晓玲;窦忠英
单  位：
河南省农业科学院畜牧兽医研究所;西北农林科技大学陕西省干细胞工程技术研究中心;宁夏医科大学总医院;西北农林科技大学西安宠物医院
关键词：
诱导性多能干细胞;细胞分化;心肌细胞;牛
摘  要：
【目的】研究牛诱导性多能干细胞(Bovine induced pluripotent stem cells,biPSCs)向心肌细胞的分化能力。【方法】将biPSCs悬浮培养制备类胚体,采用类胚体介导体外自由分化与定向诱导分化(添加RA、DMSO和RA+DMSO3种诱导物)的方法,使biPSCs在体外分化,比较了biPSCs在3种不同心肌诱导条件下定向分化为心肌细胞的诱导效率;分别提取biPSCs、类胚体及不同诱导体系分化细胞的总RNA,用RT-PCR检测心肌细胞发育标志基因的表达。【结果】biPSCs在悬浮条件下培养7d,能够形成典型球形和囊腔样类胚体。类胚体再贴壁培养7d,经过免疫细胞化学检测,贴壁细胞大量分化为α-actin阳性的心肌样细胞。在体外诱导分化体系中,RA+DMSO共同诱导的效率较RA和DMSO单独诱导显著增高(P<0.05)。RT-PCR结果显示,各组分化后的细胞中心肌细胞发育特异性基因ACTA2与GATA4均有高水平表达。【结论】biPSCs在体外悬浮培养能够形成类胚体,类胚体贴壁培养后可以分化为α-actin阳性的心肌样细胞;分化形成的心肌样细胞均表达心肌特异性基因。
译  名：
Induced differentiation of cardiomyocytes from bovine induced pluripotent stem cells
作  者：
CHEN Dong-mei1,LChang-rong1,XIN Xiao-ling1,2,DOU Zhong-ying1 (1 College of Veterinary Medicine,Northwest A&F University,Shaanxi Key Laboratory of Molecular Biology for Agriculture, Shaanxi Branch of National Stem Cell Engineering &Technology Center,Yangling,Shaanxi 712100,China;2 Institute of Animal Husbandry and Veterinary Medicine,Henan Academy of Agricultural Sciences,Zhengzhou,Henan450002,China)
关键词：
induced pluripotent stem cells(iPSCs);differentiation potential;cardiomyocytes;bovine
摘  要：
【Objective】The first evidence that bovine iPS cells could be induced to differentiate into cardiomyocytes by RA and DMSOin vitro was reported.【Method】The biPSCs were cultured using the hanging drop and suspension culture method for embryoid body(EBs)formation.Incubation of EBs with RA/MSO and RA+DMSO was used to induce cardiac differentiation.The marker proteins of cardiomyocytes was examined by immunocytochemistry staining.The induction efficiency was compared in three different cardiac induced conditions.The marker genes of cardiomyocytes including GATA4and ACTA2were exam- ined by reverse transcription polymerase chain reaction(RT-PCR).【Result】biPSCs could formate EBs af- ter 7days in suspension culture.A maximum level of spontaneously differentiation of cardiac cell clusters was observed within the outgrowths of embryoid bodies(EBs)after 7days in adherent culture.Induction with both RA and DMSO resulted in a significantly higher elevation in differentiation rate than RA or DM- SO alone(P<0.05).RT-PCR confirmed that the differentiated cells derived from biPSCs could express cardiomyocytes marker genes GATA4and ACTA2.【Conclusion】The results revealed that the established biPSCs can effectively differentiate into cardiomyocyte-like cells.Cardiomyocytes were evident as they expressed cardiac cell specific genes and protein markers including GATA4,ACTA2and alpha-actin when analyzed by PT-PCR and immunohistochemical staining.

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