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时间:2019-03-05
《大鼠骨髓间充质干细胞多向分化潜能的研究》由会员上传分享,免费在线阅读,更多相关内容在教育资源-天天文库。
1、万方数据;:。善d曼蓄煮j专羔峰三:川,。Sep,36cs,大鼠骨髓间充质干细胞多向分化潜能的研究汪浩文1何志旭26王志华2刘俊峰2(1皖南医学院附属池州医院一池州市人民医院新生儿科安徽池州247100;2贵阳医学院干细胞研究中心一贵阳医学院附属医院儿科贵州贵阳550004)【摘要】目的分离和培养大鼠骨髓间充质干细胞(marrowmesenchymalstemcells,MSCs),并行多向分化潜能的鉴定。方法经Percoll梯度分离法获得大鼠骨髓单个核细胞。贴壁培养后,通过倒置显微镜进行细胞形态学观察。并采用流式细胞术进行
2、分析。传至第3代后,分别进行成脂肪细胞、神经元样细胞、血管内皮样细胞的诱导。并行免疫组织化学鉴定。结果骨髓问充质干细胞大小较为均匀,基本上呈梭形或星形的上皮样细胞,传代培养后的细胞体积增大,成纤维样细胞逐渐增多。经反复传代纯化的Mscs检测CD71、CD29呈阳性表达并证实骨髓间充质干细胞经诱导后分化为脂肪细胞、神经元样细胞、血管内皮样细胞。结论在体外,大鼠骨髓间充质干细胞具有多向分化潜能.可作为组织工程的种子细胞。【关键词】骨髓间充质干细胞;脂肪细胞;神经元样细胞;血管内皮样细胞【中图分类号】R446.5【文献标志码】AM
3、ultilineagedifferentiationpotentialofmarrowmesenchymalstemcellsWANGHao—wenl,HEZhi—xu2^,WANGZhi—hua2,LIUJun—fen92(1DP户口rfmP竹fo.厂NPo疗“fP,C,liz^o“Hos户if口Z.H,口竹以n打^4Pdif口ZCoZZPgP,C^iz^o“2471【)O,A行^“iProui行cP,C^i行n;2SfPmCeZZR已sP口rc^Ce竹£Pro,G“iy(【”gMPdicnZU”iz圯rs打y—D已户口,
4、t埘P九fo,PPdi口frics,TheA,filintedHospitalo,GutynngMedtcQlUniversity,G“iy口行g55(JIJlJ4,G“妇^o“Proui打c已,CIlli玎c£)【Abstract】ObjectiveToisolateandculturemesenchymalstemcells(MSCs)frombonemarrowofadultratsandtoidentifythemuItiIineagedifferentiationpotentiaJofthece¨s.MeChods
5、ThemononuclearcenswereobtainedafterPercollgradientcentrifugation.AfteradherenceMSCswerecultured,themorphologicchangesofMSCsweredetectedbyinvertedlightmicroscopy.TheexpressionsofCD71,CD29,CD04andCDl45inMSCsweredetectedandjdentifjedbyflowcytometry.Thethirdpassagecell
6、swereharvestedandinducedtodifferentiateintoadiposecells,neuron—likecells,andvascularendothelium—likeceUs,thentheywereidentifiedbyimmunohistochemicalstainingrespectively.ResuItsTheresultsshowedthattheculturedMSCsweregenerallyspindle—shapedorstar—shapedepidermal一“ked
7、cells.ThecellularvolumeincreasedandfibrobIast—likeceIlswereincreasedafterserialsubcultivation.AfterseveralgenerationstheyturnedouttobepositiveexpressionsofCD29,CD71.Afterinduction,theywerecon“rmedtobeadiposecells,neuron—likecellsandvascularendotheliumlikecellsbyimm
8、unohistochemicalstaining.ConcIusionsf力口ifrDMSCshavemultilineagedifferentiationpotentialandcanbeusedastheseedcellsoftissueengineering.【Keywords】ma
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