资源描述：
《caco-细胞单层模型的建立与验证》由会员上传分享，免费在线阅读，更多相关内容在教育资源-天天文库。
1、Caco-2细胞单层模型的建立与验证Caco-2细胞单层模型的建立与验证荆,敏捷,盛星,胡一桥(南京大学医药生物技术国家重点实验室,南京210093)摘要:目的研究建立Caeo-2细胞模型的方法,并确认Caeo-2细胞模型的正常生长特性,以在日常培养中进行监控.方法常规培养条件下,Caeo-2细胞以每lmL含8X104个的接种密度接种于Millieell插入式培养皿中,培养21d后用光镜,电镜,细胞密度测定等方法检查细胞的形态学及生长特点,测定碱性磷酸酶活性,跨膜电阻,荧光素钠透过量以检验细胞单层的极化现象和致密程度.结果Caeo-2细胞单层在生长10
2、d以后均匀,致密,21d后微绒毛,碱性磷酸酶,紧密连接等呈不对称分布,跨膜电阻达到600Q-cm2,荧光素钠透过量为4.16Pg?h-em一,而同样条件下空白膜的透过量大于240pg?h一?cm-.:结论本培养条件下,Caeo-2细胞单层形态与小肠上皮细胞类似,跨膜电阻,标志物透过量均达到要求,可以作为研究小肠药物转运过程的体外模型.关键词:Caeo-2细胞;转运;跨膜电阻;渗透性中图分类号:Q81l文献标识码:A文章编号:1001—2494(2006)18—1431—04EstablishmentandValidationofCaco-2CellLi
3、nesforLntestinalEpithelialPemeabilitySUNMin-jie,SHENGXing,HUYi-qiao(StateKeyLaboratoryofPharmaceuticalBiotechology,NanjingUniversity,柳ng210093,Ch/na)A】BsTRACT:o]BJE(1,,EToinvestigatetheestablishmentofCaeo-2cellmonolayermode1.andtoidentifyCaeo-2'Snaturalgrowthinor.dertomonitorthe
4、dailycultivation.METHODSCaeo-2cellwascultivatedinMillieellplateinsertswith8X104cells?mL—lcelldensityinstandardproceduresfor21d.rhemorphologicalandgrowthcharactersofthemonolayerwereexaminedwithopticalmicroscope.electronmi.eroscopeandcelldensitydetemfination.111earizationandeompae
5、tifieationofthemonolayerWasdeterminedbythealkalinephosphataseac—tivity,transepithelialeleetrica]resistance(TEER)andapical—to-basolateralofsodiumlluoresceinacrosscellmonolayem.RESULTSCaeo-2cellmonolayerbecamewell-propo~onedandcompactafter10dgrowth.After21d,miemvillus,alkalinephos
6、phataseandtightconjunctionswereasymmetricallydistributed,TEERvaluereached600Q?em_.,andthetransportationofsodiumlluoresceinWasq-.16-h一?em一transportationacrossnon—cellmembranewasmoilthan240pg?h?eminthesameeonditions.CONCLUSIONInthesecultivationeondi—tions,theCaeo-2cellmonolayerhad
7、becomeananalogyofsmallintestinalepithelium.ThevalueofTEERandtransportationofsodiumlluoils—eeinallreachedthecriterions,whichmeantthatthemonolayerbecameanvaluablemodelsystemforintestinalepithelialpermeability.Inthisresearch,Caeo-2cellmonolayermodelWasestablishedandevaluated.旺YWORI
8、~:Caeo-2cell;transport;transepithelialelectrica
