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ID:31072282
大小:2.97 MB
页数:6页
时间:2019-01-06
《脐血间充质干细胞分离,培养与鉴定》由会员上传分享,免费在线阅读,更多相关内容在工程资料-天天文库。
1、脐血间充质干细胞分离、培养与鉴定李启明1程天民1李宁2*通信作者粟永萍1冉新泽1(1.第三军医大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,高压氧中心重庆400038)摘要:目的研究脐带血(HUCB)中含有的间允质干细胞(MSCs)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定获得的MSCs结果来源于脐血的单个核细胞经体外培养
2、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、培养出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-ping,RUANxin-ze(1.StateKeyLaboratoryofTrauma,BurnsandCombi
3、nedInjury,InstituteofCombinedInjury,2.HyperbaricoxygenCenterXinqiaoHospital,ThirdMilitaryMedicalUniversity,Chongqing,400038,China)Abstract:ObjectiveToinvestigatetheisolation,cultivation,differentiationandidentificationofhumanumbilicalcordblood(HUCB)mesenchymalstemcells(MSCs)invitro,andtoobservethef
4、easibilityofusingMSCsasseedcellsinexperimentalfurthermore.MethodsHUCBwerecollectedfromfulltermchildbirthscheduledforCesareansectionandvaginadeliveries,a11specimenswasobtainedsterilelywithPreservative-freeCPDA-1compoundnatriumcitricum.Afterlyseredbloodcell,thecordbloodmononuclearcellwasisolatedbyhum
5、anlymphocyteseparatingmedium.culturewithdulbecco'smodifiedeagle'smedium(DMEM/F12)mediumtoproduceadherentlayerandidentificationwithorientationdifferentiationresults.ResultsTheUCB-derivedmononuclearcells,whensetinculture,gaverisetoadherentcells,whichexhibitedeitherafibroblast-1ikebutcrescent-shapedor
6、bending-shapedmorphologyormesenchymal-likephenotype.ItcanbeaffirmedastheMSCswhichcharacterizedwithmulti-differentiatingpotentialityafterthemyogenic、lipogenicandneurogenicdifferentiation.ConclusionMSCsinHUCBcanbeisolatedandcultivatedinvitro,whichcouldberegardedasanalternativesourceofMSCsforfurtherex
7、perimentaltests。Keywords:umbilicalcordBlood;mesenchymalstemcells;cultureinvitro;differentiationandidentification目前认为,成体干细胞实际上主要是指在不同组织中的间充质干细胞,(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
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