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1、脐血间充质干细胞分离、培养与鉴定李启明I程天民I李宁"粟永萍I冉新泽1(1.第三军医人学全军复合伤研究所，创伤、烧伤与复合伤国家重点实验室2.第三军医大学新桥医院，高压氧中心逼庆400038)摘要：目的研究脐带血(HUCB)中含有的间允质干细胞(1SCs)体外分离、培养条件，探讨其作为种子细胞用于下一步实验研究的稳定性与实用性。方法无菌条件下取正常足生产的脐带血，经CPDA-1复合抗凝剂抗凝，然后经过去红细胞处理，再用淋巴细胞分离液分离脐血的单个核细胞，DMEM/F12培养基进行培养和纯化获得贴壁细胞层，采用定向诱导分化方法检测鉴
2、定获得的MSCs结果来源于脐血的单个核细胞经体外培养贴壁后出现形态学上可见间充质样的细胞，问充质样细胞为类似成纤维样的细胞形态，但偏向半月形弯曲状，经过成肌、成脂肪、成神经细胞诱导分化成功后确定为间充质干细胞“结论脐血来源的间充质干细胞能够在体外分离、培养岀有用的进一步实验所需要的种子细胞。关键词：脐带血；间充质干细胞；细胞培养；诱导分化鉴定中图法分类号：文献标识码：THEISOLATIONCULTUREANDIDENTIFICATIONOFHUCAMSCsLIQi-ming,CHENTian-minLIning,SUYong-pi
3、ng,RUANxin-ze(1.StateKeyLaboratoryofTrauma,BurnsandCombinedInjury,InstituteofCombinedInjury,2.HyperbaricoxygenCenterXinqiaoHospital,ThirdMilitaiyMedicalUniversity,Chongqing,40003&China)Abstract:ObjectiveToinvestigatetheisolation,cultivation,differentiationandidentifica
4、tionofhumanumbilicalcordblood(HUCB)mesenchymalstemcells(MSCs)invitro,andtoobservethefeasibilityofusingMSCsasseedcellsinexperimentalfurthermore.MethodsHUCBwerecollectedfromfulltermchildbirthscheduledforCesareansectionandvaginadeliveries,al1specimenswasobtainedsterilelyw
5、ithPreservative-freeCPDA-1compoundnatriumcitricum・Afterlyseredbloodcell,thecordbloodmononuclearcellwasisolatedbyhumanlymphocyteseparatingmedium,culturewithdulbecco'smodifiedeagle'smedium(DMEM/Fl2)mediumtoproduceadherentlayerandidentificationwithorientationdifferentiati
6、onresults.ResultsTheUCB-derivedmononuclearcells,whensetinculture,gaverisetoadherentcells,whichexhibitedeitherafibroblast-likebutcrescent-shapedorbending-shapedmorphologyormesenchymal-likephenotype.ItcanbeaffirmedastheMSCswhichcharacterizedwithmulti-differentiatingpoten
7、tialityafterthemyogenic>lipogenicandneurogenicdifferentiation.ConclusionMSCsinHUCBcanbeisolatedandcultivatedinvitro,whichcouldberegardedasanalternativesourceofMSCsforfurtherexperimentaltestsoKeywords:umbilicalcordBlood；mesenchymalstemcells；cultureinvitro；differentiatio
8、nandidentification通信作者目前认为，成体干细胞实际上主要是指在不同组织屮的间充质干细胞,（MSCs）,它是广泛存在生物体尤其骨髓内的一类干细胞，以前被称为基质干细胞，它最初是由Fnedemteinr发现的易于贴附于塑
