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时间:2020-04-19
《人卵巢癌组织肿瘤干细胞的分离和鉴定.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
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:
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