无血清培养法分选舌癌侧群细胞及其特性的鉴定

《无血清培养法分选舌癌侧群细胞及其特性的鉴定》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、无血清培养法分选舌癌侧群细胞及其特性的鉴定王景涛，季平，邓诚，孙守娟，邹波（400015重庆，重庆医科大学附属口腔医院口腔颌面外科）[摘要]目的探索人舌鳞癌细胞株Tca8113中侧群细胞的分选方法，以期获得较好的富集分选侧群细胞的方法。方法利用流式细胞仪分选传统含血清培养方法获得的人舌鳞癌细胞系Tca8113中的侧群细胞（SP），并与无血清培养方法分离出的侧群细胞（TSC-SP）进行形态学比较和平板形成克隆实验及成瘤实验的比较，富集并验证肿瘤干细胞相关亚群。结果人舌癌细胞系Tca8113中SP细胞的含量为0.2﹪，而Tca811

2、3细胞系的细胞在无血清培养基中能够成活，3-5d后增殖并形成悬浮的肿瘤干细胞球，并随着培养时间的延长而肿瘤干细胞球体积增大和更加致密；而应用无血清悬浮法培养的Tca8113细胞系中TSC-SP细胞的含量为0.4﹪；SP与TSC-SP细胞在平板形成克隆实验的形成率均为100﹪；接种16d后，104、105数量级的TSC-SP和SP接种裸鼠背部均出现瘤体。结论人舌鳞癌细胞系Tca8113中含极少量的SP细胞，利用无血清培养基可以达到富集肿瘤干细胞亚群的目的。[关键词]人舌鳞癌；无血清培养；肿瘤干细胞；侧群细胞；流式细胞仪[中图法分类

3、号][文献标志码]AIsolationandidentificationofsidepopulationcellsfromhumantonguesquamouscarcinomaTca8113celllineunderserum-freecultureconditionWangJingtao,JiPing,Dengcheng,SunShou-uan，ZouBo(DepartmentofDentofacialSurgery,theAffiliatedStomatologyHospital,ChongqingMedicalUnive

4、rsity,Chongqing,400015)[Abstract]ObjectiveToinvestigatesidepopulation(SP)cellsinhumantonguesquamouscarcinomacelllineTca8113underserum-freeculturemedium(SFM)orserum-supplementedmedium(SSM)，andtodetectproliferativeanddifferentialability,tumorigenicabilityofSPcells，andt

5、oidentifytumor-likestemcellsinTca8113,thepresentstudyisdesigned.MethodsInvitroindividualcellculturewasemployedtoobservetheproliferatingcharacterofTca8113.SPweresortedandcomparedbyflowcytometry.Tca8113celllinewascultivatedinSFMandthecancerstem-likecellsreformingintofl

6、oatingsphereswereisolated,wasnamedTSC-SP.TheproliferativeabilityofSPcellsinvitrowasdetectedbyself-renewalassays.Thehetero-transplanttumormoldwanestablishedusingnudemicebysubcutaneousinjectionoftumorcells.Thetumorigenicpropertiesofthedifferentclusterswereinvestigatedi

7、nvivo.ResultsSPwaspositivelyexpressedonly0.2%ofthecellsinhumantonguesquamouscarcinomacelllineTca8113,butTSC-SPwas0.4%;ComparedwithTSC-SPandSP，TSC-SPcellsshowedincreasedproliferativecapacity.SPandTSC-SPcellswerepossessedproliferative,self-renewalanddifferentiationpote

8、ntialwhichwereresponsibleforthefloatingtumorclone.TheTSC-SPcellswerepossessedmoreclonogenicandinnudemice,only104SPcellsandTSC-SPcel