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1、乳腺癌新辅助化疗后的癌干细胞分离培养董华英，吴诚义，陈元文(400016重庆，重庆医科大学附属第一医院内分泌外科)[摘要]目的：从不同周期新辅助化疗后的乳腺癌组织屮分离培养乳腺癌干细胞(breastcancerstemcells,BCSCs),观察其体外增殖和分化特性，研究新辅助化疗对BCSCs分离培养的影响。方法：在添加生长因子的无血清培养基(serumfreemedia,SFM)中悬浮培养细胞获得富含BCSCs的微球体(nianunospheres,MSs),通过克隆形成实验研究微球体细胞(mammos
2、pheres-derivedcells,MSDCs)的自我更新能力:在添加5%胎牛血清的培养基中培养MSDCs诱导其分化,研究其分化能力;免疫细胞化学检测ESA和Nestin在MSDCs中的表达;流式细胞术(FlowCytometry,FCM)检测MSDCs屮的ALD111•细胞含量。结果：化疗1个周期的5例乳腺癌标本经培养均无MSs形成，化疗2个周期的6例标本经培养其中3例有MSs形成,化疗3个周期以上的9例标本中的5例可以直接分离出MSs,其余4例仅得到少量坏死的细胞和组织碎片：MSDCs在含血清的培养
3、基屮贴堆分化：MSDCs均含有ALDII1•细胞。结论：从新辅助化疗后的乳腺癌组织中分离BCSCs宜选取化疗两个周期后的标本；新辅助化疗可以富集BCSCs；ALDlir表型的BCSCs比CD14CD24表型的BCSCs在乳腺癌屮分布更加广泛。[关键词]乳腺肿瘤;化疗；肿瘤干细胞:细胞分离:细胞培养IsolationandcultureofbreastcancersteincellsderivedfrombreastcancerafterneoadjuvantchemotherapyDongHuaying,W
4、uChengyi,ChenYuanwen(DepartmentofEndocrineSurgery,FirstAffiliatedHospital,ChongqingMedicalUniversity,Chongqing,400016,China)[Abstract]Objective:Toisolateandcultureofbreastcancerstemcells(BCSCs)frombreastcancertissuesafte『differentcyclesofneoadjuvantchemoth
5、erapy,andinvestigatetheirproliferationanddifferentiationbiologicalcharacteristicsinvitroinordertoobtaintheimpactofneoadjuvantchemolherapyontheisolationandcultureofBCSCs・Methods:Breasttissuesamplesfrom20patientsreceivingTACneoadjuvantchemotherapyinourdepart
6、mentfromOctober2009toApril2010werecollected・Therewere5caseswith1cycleofchemotherapy.6caseswith2cycles,7caseswith3cycles,and2caseswith4cycles・Thetissuesamples(1cmxlcmxlcm)wereobtainedthroughoperationandcutintopiecesforculture・Thesuspendedcellswereculturedin
7、serum-freemedia(SFM)supplementedwithgrowthfactors.Cellsgrownintheseconditionsasnonadherentsphericalclustersofcells(usuallynamed"mammospheres'；MSs)wereenrichedinBCSCs.Thesell-renewalcapacityofmaininospheres-derivedcells(MSDCs)wasevaluatedbycolonyformationas
8、say・Conversely,thedifferentiationofMSDCswasinducedbyculturingtheminmediasupplementedwith5%fetalbovineserumw让houtgrowthfactors;Expressionsofepithelial(ESA)andNestininMSDCswasdetectedbyimmunocytochemistry.ALDH1
