Caco_2细胞单层模型的建立与验证.pdf

Caco2 * 孙敏捷, 盛星, 胡 一桥 ( , 210093) : 研究建立Caco2 细胞模型的方法, 并确认Caco2 细胞模型的正常生长特性, 以在日常 养中进行监控 4 常规 养条件下, Caco2 细胞以每 1 m 含8 10 个的接种密度接种于Millicell 插入式 养皿中, 养21 d 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 Caco2 细胞单层在生长 10 d 以后均匀致密, 21 d 后微绒毛碱性磷酸酶紧密连接等呈不对称分布, 2 - 1 - 2 - 1 - 2 跨膜电阻达到600 cm , 荧光素钠透过量为416 gh cm , 而同样条件下空白膜的透过量大于240 gh cm 本 养条件下, Caco2 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 : Caco2 细胞; 转运; 跨膜电阻; 渗透性 : Q811 :A : 1001- 2494( 2006) 18- 1431- 04 Establishment and Validation of Caco2 Cell Lines for Lntestinal Epithelial Permeability * SUN Minjie, SHENG Xing,HU Yiqiao ( StateKey Laboratory of Pharmaceutical Biotechology, Nanj ing University, Nanj ing 210093, China) ABSTRACT: OBJECT VE To investigate the establishment of Caco2 cell monolayer model, andto identify Caco2s natural growth in or 4 - 1 der to monitor the daily cultivationMETHODS Caco2 cell was cultivated in Millicell plate inserts with 8 10 cellsm cell density in standard procedures for 21 dThe morphological and growth characters of the monolayer were examinedwith optical microscope, electron mi croscope and cell density determinationThe polarization and compactification of the monolayer was determined by the alkaline phosphatase ac tivity, transepithelial electrical resistance(TEER) and apicaltobasolateral of sodium fluorescein across cell monolayersRESULTS Caco2 cell monolayer became wellproportioned and compact after 10 d growthAfter 21 d, microvillus, alkaline phosphatase and tight conjunctions - 2