人卵巢癌组织肿瘤干细胞的分离和鉴定.pdf

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1、人卵巢癌组织肿瘤干细胞的分离和鉴定林盛龙海霞向橦朱波谢荣凯[摘要]目的从人卵巢癌组织中分离并鉴定卵巢癌干细胞。方法取新鲜卵巢癌组织，以胶原酶和透明质酸酶充分消化，获得单细胞悬液，经红细胞裂解液处理，CD133磁珠孵育进行磁性细胞分选，流式细胞仪验证分选效率。将阳性细胞置于含人表皮生长因子、碱性成纤维细胞生长因子、牛血清白蛋白和人胰岛素的无血清培养基中，待细胞成球状生长后进行免疫荧光、分化实验和成瘤实验，鉴定其干细胞特性。结果从人卵巢癌组织中成功分离出肿瘤干细胞，流式细胞仪检测其CD133的表达率为82.5％。该细胞在无血清培养基中呈悬浮球状生长，具有自我更新能力、分

2、化潜能和更强的致瘤能力，具备干细胞特性。结论人卵巢癌组织中存在卵巢癌干细胞，利用CD133磁珠分选得到的肿瘤干细胞具备干细胞的相关特性，可用于后续卵巢癌干细胞生物学特性的研究。肿瘤干细胞；卵巢肿瘤；细胞分离IsolationandidentificationofcancerstemcellsfromprimaryhumanovariancancertissuesLINShengLONGHai-xiaXIANGTongZHUBoXIERong-kaiDepartmentofOncology,XinqiaoHospitaloftheThirdMilitaryMedica

3、lUniversity,Chongqin400038,China[Abstract]ObjectiveToisolateandidentifythecancerstemcellsfromprimaryhumanovariancancertissues.MethodsFreshtumortissuesfromfivecasesofpathologicallydiagnosedovariancancersweretaken,mincedandthendigestedwithcollagenaseandhyaluronidasetoobtainsinglecellsusp

4、ension.TheerythrocyteswereremovedwithACKLysisbuffer.Thesuspensionsweresortedbymagneticactivatedcellsorting(MACS)usingCD133-bindingmicrobeads.ThenthesortedCD133+cellswereverifiedbyflowcytometry.Thecellswereculturedinserum-freemediumsupplementedwithEGF,bFGF,insulinandBSA,andgrewintospher

5、oids.Immunofluorescence,differentiationandtumorformationtestsofthecellswereperformedtocharacterizethepropertiesofcancerstemcells.ResultsTheovariancancerstemcellsweresuccessfullyisolatedfromprimaryhumanovariantumors,whichformedtypicalspheroidsinserum-freemediumandhadstrongerabilityoftum

6、origenesis.TheresultsofrelatedexperimentsverifiedthatCD133positivecellsownedthepropertiesofcancerstemcells.ConclusionsTheovariancancerstemcellspresentingthecharacteristicsofsternnessinvitroandinvivo,havebeensuccessfullyisolatedfromprimaryhumanovariantumortissuesbyMACS.Theisolatedovaria

7、ncancerstemcellscouldbeusedinfutureresearchesoftheirbiologicalfunctions.Tumorstemcells;Ovarianneoplasms;Cellseparation10.3760/cma.i.issn.0253-3766.2011.12.004400038重庆，第三军医大学附属新桥医院全军肿瘤研究所[林盛（现工作单位：646000泸州医学院附属医院肿瘤科）、龙海霞、向橦、朱波]妇产科（谢荣凯）·897··898·@@[1]陈运贤，钟雪云.白血病干细胞的靶向治疗.中华肿瘤杂志，2006,28: