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时间:2017-12-07
《人脐血间充质干细胞源性心肌细胞的诱导分化研究》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、国际心血管病杂志2010年9月第37堂箜塑!!塑!!兰望!!竺竺:!:!!!:·315·人脐血间充质干细胞源性心肌细胞的诱导分化研究杨水祥黄景玲【摘要】目的:探讨体外人脐血间充质干细胞(hMSCs)诱导分化成心肌细胞的可行性和最佳方法。方法:收集健康产妇脐血细胞,分离单个核细胞,从中进一步分离间充质干细胞,传代培养至第三代,应用免疫荧光流式细胞仪标记MSCs特异性抗原CD34,CD44和CD90。5-氮(杂)胞苷(5一aza)诱导分化4周后,免疫组织化学染色和RTPCR分别检测心肌细胞标志物肌钙蛋白I,GATA4和p一肌球蛋
2、白重链。结果:脐血源性MSCs经5-aza诱导分化后,呈现成纤维细胞样形态和克隆增殖特点。免疫分型与骨髓来源MSCs一致,且免疫组织化学染色和RT-PCR可检测到肌钙蛋白I、GATA4和一肌球蛋白重链的表达。结论:脐血源性MSCs能够被诱导分化成心肌样细胞,可成为干细胞移植的细胞来源。【关键词】间充质干细胞;脐血细胞;5-氮(杂)胞苷;心肌细胞DOI:10.3969/j.issn.1673—6583.2010.04.019HumanMSCsfromUCBcanbeintroducedtodifferentiateintoca
3、rdiomyocytesinvitroYangShui—xiangHuangJing—ling.1.DepartmentofCardiology,BeijingShijitanHospital,Beijing100038,China;2.DepartmentofCardiology,QinghaiProvinceCardiovascularFacultyHospital,Xining,Qinghai810012,China[Abstract]Objective:Inordertodetectthepossibilityand
4、thebestwayformesenchymalsterncells(MSCs)fromumbilicaIcordblood(UCB)tobeintroduceddifferentiati0nintocardiomyocytesinvitro.Methods:Theumbilicalcordbloodwascollectedfromthehealthdeliverywomenwithinformedcon—sent,inwhichthemononuclearcellswereisolated,andMSCswerefurth
5、erseparated,expandedtothe3passage.ThecharacteristicmarkerofMSCsofCD34,CD44andCD90wasidentifiedwiththeflowcytometryapplyingimmunofluorescenceway.Followingdifferentiationintroducedwith5-azafor4weeks,themyocardialmarkersofTroponinI,GATA4and8一MHCweredetectedbyimmunohis
6、tochem—istrystainingandRT—PCR,respectively.Results:TheMSCsinducingwith5-azashowedchangesoffi—broblast—likemorphologyandclonallyexpandedcharacteristics.Theimmunophenotypeofthesecellswasconsistentwiththatreportedfrombonemarrowmesenchymalstemcells.Themyocardialmarkers
7、ofTroponinI,GATA4andMHCwerepositivelydetectedbyimmunohistochemistrystainingandRT—PCR,respectively.Conclusion:OurdataindicatethatumbilicaIcordbloodcanbeservedasavaluableandconvenientSOUrCeofmesenchyma1stemcells.whichcandifferentiateintocardiomyocyte-likecell。andcanb
8、eusedasanalternativesourceinplaceofbonemarrow-derivedstemcells.[Keywords]Mesenchymalstemcells;Umbilicalcordblood;5-aza;Cardiomyocyte越来越多的证据表明,成人来
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