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1、与胎儿心肌细胞共同培养诱导人胚胎干细胞发生心肌样分化作者：常静1,雷寒1,陈建斌2,贾锋鹏1单位：重庆医科大学附属第一医院：1心血管内科,2血液内科，重庆400016提要：目的了解与胎儿心肌细胞共同培养对人胚胎干细胞(embryonicstemcells,ESCs)向心肌方向分化的影响。方法通过酶消化法从流产胎儿中分离培养出胎儿心肌细胞及成纤维细胞,将后者用丝裂霉素处理后制备成滋养层；收集辅助生育屮心施行试管婴儿后所剩余的健康胚胎，取内细胞团接种在滋养层上分离培养岀人ESCso用荧光原位杂交(fluor
2、escentinsituhybridzation,FISH)方法來检测上述两种细胞的XY染色体，选用不同性别的胎儿心肌细胞与2~5代人ESCs以1：2的比例混合,然后以5×104/cm2的密度接种于培养皿屮进行共同培养，诱导其向心肌方向的分化。将未与胎儿心肌细胞混合的人ESCs以相同密度接种于培养皿中作为阴性对照。至第5天收集共同培养的细胞,进行双重染色，即FISH染色及心肌特异性蛋白Desmin或cTnl免疫组化染色。结果在与胎儿心肌细胞共同培养5d后，40%~50%的人ESCs在心肌细胞
3、的诱导下表达心肌特异性蛋白Desmin或cTnl,而没有进行共同培养的人ESCs不表达上述蛋白。结论与胎儿心肌细胞的共同培养能诱导人ESCs发生向心肌方向的分化，人ESCs有望成为心肌干细胞移植的细胞材料。关键词:胎儿心肌细胞;人胚胎干细胞;共同培养;心肌样分化中图法分类号:R322.11;R329.21文献标识码:ACardiacdifferentiationofhumanembryonicstemcellsbycoculturingwithhumanfetaicardiomyocytesCHANGJ
4、ingl,LEIHanl,CHENJian~bin2,JIAFeng~pengl(lDepartmentofCardiology,2DepartmentofHemotology,TheFirstAffiliatedHospitalofChongqingMedicalUniversity,Chongqing400016,China)AbstractObjectiveToexplorethecardiacdifferentiationofhumanembryonicstemcells(ESCs)incocu
5、lturingwithhumanfetalcardiomyocytes.MethodsFetalcardiomyocytesandibroblastswereobtainedfromtheabortedfetus,andhumanESCswerefromtheinnercellmass(ICM)ofthesurplusembryosfortubebabies,withthetreatedfibroblastasfeederlayer・HumanESCsofpassa^e2to5werecocutured
6、withfetalcardiomyocytesattheratioof1:2,andtheXYchromosometypesofthesetwokindsofhumancellsweredifferenttoeachotherinordertobemarked.Atthesametime,humanESCswithoutcoculturewereusedasnegativecontro.1Thecoculturedcellswereharvested5dayslaterandweredouble-sta
7、inedwithhumanXYchromosomeprobeandcardiacspecificantibody(desminorcardiactroponinI,cTnI).ResultsOncocultureday5,40%~50%humanESCsexpressedcardiacspecificantigens,andhumanESCswithoutcoculturedidnotexpressthoseantigens・ConclusionHumanESCscouldbeinducedintoca
8、rdiacdifferentiationbythecoculturewithfetalcardiomyocytes,andhopefullytheywillbethecandidateforcelltransplantationofhear・tKeywords:fetalcardiomyocyte;humanembryonicstemcells;coculture;cardiacdifferentiation心血管疾病已经成为了工业化社会屮
