人脐带间充质干细胞体外分离、纯化及鉴定

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1、暨南大学学报(医学版).JournalofJinanUniversity(MedicineEdition)人脐带间充质干细胞体外分离、纯化及鉴定王娟，陆琰，何冬梅，张疽(庇南大学医学院血液病研究所，广东广州510632)【摘要］目的：从人脐带中分离、培养并鉴定间充质干细胞(MSCS)O方法:用胶原酶消化法分离脐带间充质干细胞，差速贴壁法进行纯化;MTT法检测细胞增殖活性，并绘制生长曲线;流式细胞仪检测其表面标志和细胞周期;地塞米松、抗坏血酸、磷酸甘油诱导其向成骨细胞分化，并用碱性磷酸酸染色鉴定;地塞米松、胰岛索、号I喙美辛诱导其向脂肪细胞分化，用油红0染色鉴定;将脐带MSCs注射到BAL

2、B/c裸鼠肾被膜下，观察有无致瘤性。结果:从人脐带中分离出的间充质干细胞为梭形，呈平行排列生长或漩涡状生长;细胞表达CD29.CD44,低表达CD106,不表达CD14、CD31、CD34、CD45和HLA-DR；具有分化成脂肪细胞和成骨细胞的潜能;将脐带MSCs移植到BALB/c裸鼠肾被膜下，未观察到致瘤性。结论:从脐带中成功分离培养的细胞，具有间充质干细胞生物学待性。［关键词］脐带；间充质干细胞；分离；细胞培养；鉴定［中图分类号］Q813［文献标识码］AJ文章编号］1000-9965(2009)04-0367-06Isolation,purificationandidentifica

3、tionofmesenchymalstemcellsderivedfromhumanumbilicalcordWANGJuan,LUYan,HEDong-mei,ZHANGYuan(InsitituteofHematology,MedicalCollege,JinanUniversity,Guangzhou510632,China)2008-12-02［收稿日期］［基金项目］［作者简介］[Abstract]Aim：Toisolate9purifyandidentifymesenchymalstemcellsderivedfromhumanumbilicalcord.Methods:Mes

4、enchymalstemcellswereisolatedfromhumanumbilicalcordbydigestionofcolla・genaseIVandtheiruniquegrowthcharacteristicofattachingtothewallofcellcultureflask.ThecellproliferationwasanalyzedbyMTTassayandthegrowthcurvewasdrawn.Theirsurfacemarkersweredetectedbyflowcytometry.Thecellswereinducedtoosteoblast

5、withdexamethasone,antiscorbicacidandp-sodiumglycerophosphate.Andtheyalsowereinducedtoadipocyteswithdexamethasonefinsulinandindomethacin.Afterinduction9thecellswereobservedbyALPstainingandoilred0staining.TheumbilicalcordMSCswasinjectedintotheBALB/cnudemicekidney'scapsuletoobservetumorigenicity・Re

6、sults:Mesenchymalstemcellsderivedfromhumanumbilicalcordwerespindle-shapedadherentcells.Theyhavestrongproliferationability.IlieywerestronglypositiveexpressionforCD44andCD29.lessexpressionforCD106,butnegativeexpressionforCD14,CD31,CD34,CD45andHLA-DR.Afterinduction,oilred0stainingandALPstainingwere

7、positive・Aftertransplantationtwefindnotumori-genicity.Conclusion：Theadherentcellsisolatedfromfromtheumbilicalcordhavebiologicalcharacter-国务院侨办重点学科建设基金项目(51205002)王姐(1983-),女■硕士研究生，研究方向：干址胞分化研究通讯作者：何冬梅，女.副研究员，硕士生导师,Tel：