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《Journal of Xi'an Jiaotong University(Medical Sciences)》 2017-02
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Inducing bone marrow mesenchymal stem cells to differentiate into cardiomyocyte-like cells via Wnt-11 in vitro

LIU Yuan;WANG Hai-ping;L Yang;WANG Hao-yu;HUO Yan-li;Department of Histology and Embryology,Hebei North University;Department of Human Anatomy,Hebei North University;  
Objective To probe into the optimal concentration of Wnt-11 to induce the differentiation of bone marrow mesenchymal stem cells(BMMSCs)into cardiomyocyte-like cells in vitro.Methods BMMSCs were isolated from the bone marrow of SD rats using whole bone marrow culture method.After cultured for 48 h,BMMSCs of the second generation were utilized for directed induction.Based on the final concentration of Wnt-11,BMMSCs were divided up into Group A(100 ng/mL),Group B(200 ng/mL),Group C(400 ng/mL)and Group D(blank control).After 72-hour induction,the cells were cultured in complete medium for 4 weeks while cells in Group D were cultured only in the complete medium.The morphological changes were observed under the phase contrast microscope.Surface antigen expression of BMMSCs was identified by flow cytometry.When cells were cultured for 4 weeks,the expressions of Desmin,Connexin43 and cTnI were detected by immunocytochemistry.Meanwhile,the ultrastructural changes were observed using transmission electron microscope. The mRNA expressions of cardiac transcription factors GATA-4,Nkx2.5 andα-MHC in BMMSCs were detected by RT-qPCR at1,2 and 4 weeks after induction.Results Primary BMMSCs formed cell colonies at 2 weeks;the cells were mainly fusiform or star-shape,and a few irregularly-shaped ones were also visible.The passaged cells were larger than those of primary culture.After induction,the cells exhibited long shuttle-shape and were aligned in parallel.Flow cytometery displayed that the positive rate of the surface antigens of BMMSCs CD29,CD45,and CD90 was97.9%,0.4% and 99.5%,respectively.When BMMSCs-induced via Wnt-11 were cultured for 4 weeks,Desmin,cTnI and Connexin43 were all positively expressed in induction groups.Whereas in the blank control group they were slightly positive or negative;the positive rate in Group B was the highest(P0.05).Transmission electron microscopy exhibited that organelles such as rough endoplasmic reticulum,mitochondria,as well as some ribosomes were visible in the cytoplasm of these cells in each induction group.In addition,myofilaments were arranged in parallel in the cytoplasm.The cells in induction groups could express GATA-4 and Nkx2.5 in the first week,and then the expression of them decreased in the second week,but then increased in the fourth week;gene expression in induction Group B was significantly higher than in the other two induction groups(P0.05).The expression of GATA-4 and Nkx2.5 in Group D was 1,α-MHC was not expressed in the four groups during the culture period.Conclusion Wnt-11 can induce the differentiation of BMMSCs into cardiomyocyte-like cells in vitro,and the optimal concentration of Wnt-11 is 200 ng/mL.
【Fund】: 河北省自然科学基金(No.H2015405017);河北省自然科学基金(No.H2014405005);; 河北北方学院创新培育项目(No.CXRC1315);; 河北省教育厅重大项目(No.ZH2012001)~~
【CateGory Index】: R54
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【Citations】
Chinese Journal Full-text Database 1 Hits
1 ZHANG Yong,CHENG Jing-liang,LI Hua-li,et al(The First Affiliated Hospital of Zhengzhou University,Zhengzhou,Henan 450052,P.R.China);In vitro isolation and primary culture of bone marrow mesenchymal stem cells[J];中国医学工程;2011-01
【Co-citations】
Chinese Journal Full-text Database 5 Hits
1 Sun Yao;He Yulan;Yu Xiaolong;Cheng Yan;Song Boyi;Gerontology Department of Affiliated Hospital of Hebei United University;;Construction of vascularized tissue-engineered myocardium in vitro[J];解剖学杂志;2015-03
2 XIE Xingwen1,HOU Feiyi2,LI Ning2,et al.1 The First Affiliated Hospital of Gansu College of Traditional Chinese Medicine;2 Gansu College of Traditional Chinese Medicine,Lanzhou 730050,China;Study of different methods of labeling for rat mesenchymal stem cells[J];中国骨质疏松杂志;2013-02
3 Zheng Song-wen 1,Wang Nan 1,Wang Yan 2,Liu Yang 2,Jiang Yuan 1,Dong Yue 1 1 Department of Radiology,2 Sino-UK Regenerative Medicine Center,First Affiliated Hospital of Dalian Medical University,Dalian 116011,Liaoning Province,China;Culture of bone marrow meenchymal stem cells from a rabbit model of steroid-induced avascular necrosis of the femoral head[J];中国组织工程研究;2012-36
4 Liu Bo-wu,Lü An-lin,Huang Wei,Hou Jing,Hou Zhao-lei,Hou Hong,Da Jing,Yang Na Department of Cardiology,Xijing Hospital,Fourth Military Medical University of Chinese PLA,Xi'an 710032,Shaanxi Province,China;Induced differentiation of bone marrow mesenchymal stem cells into cardiomyocyte-like cells by four inductors[J];中国组织工程研究;2012-14
5 ZHANG Yong-yun1,PANG Rong-qing2,HE Jie2,ZHAO Jing2,QU Liang2, WANG Gui-hua2,PAN Xing-hua2,ZHANG Cheng3(1.Center for the Basic Agricultural Experiments and Teaching,Yunnan Agricultural University,Kunming,Yunnan 650201,P.R.China;2.Center for Stem Cell and Tissue & Organ Engineering of Kunming General Hospital of People's Liberation Army,Kunming,Yunan 650032,P.R.China;3.Department of Neurology,First Affiliated Hospital,Sun Yat-sen University,Guangzhou,Guangdong 510080,P.R.China);An isolation method of embryonic-like stem cells derived from adult bone marrow[J];中国现代医学杂志;2012-09
【Secondary Citations】
Chinese Journal Full-text Database 1 Hits
1 ZHOU Yong-zhi ZHANG Shi-ming LI Ming-zhong Department of Neurosurgery,The First Hospital Affiliated to Suzhou University,Suzhou 215006,china;The research of biocompatibility of bone marrow stem cells on silk fibroin films[J];中华神经外科杂志;2006-10
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