Caco-2细胞模型验证指标的选择与评判

 目的建立Caco-2细胞模型并探讨验证该模型指标的选择与评判。方法在所建立的Caco-2单层细胞模型上,通过测定跨上皮细胞膜电阻(TEER),比较膜两侧碱性磷酸酶(ALP)活性,计算几种公认的标记物在单层模型转运参数等对Caco-2细胞单层可靠性进行评价。结果TEER在细胞单层形成过程中稳定增长,膜两侧培养液中ALP活性差异逐步加大,细胞旁及跨细胞转运标记物均呈现可预见的低或高表观渗透系数(P_app)值,P-gp标准底物非索非那定双侧转运流出率(ER)大于2。结论建立的Caco-2细胞模型在完整性、细胞极性、通透性以及P-gp表达等方面均符合胃肠吸收化合物机制研究的要求。

Selection and Utilization on the Evaluation Criterions of Caco-2 Cell Model

OBJECTIVE To establish and validate Caco-2 cell monolayer model.METHODS Caco-2 cell monolayer was evaluated by determining transepithelial electrical resistance(TEER),comparing bilateral ALP activity,and transporting standard markers in model.RESULTS TEER and the difference of bilateral ALP activity were increased in cell monolayer developing process,and the apparent permeability coefficients(Papp) of the markers of paracellular and transcellular transport were all within the reported range.The efflux rate of fexofenadine(the standard substrate of P-gp) was above 2.CONCLUSION The integrity and permeability of cell monolayer,cell polarity and P-gp expression in cell model were satisfactory,which showed that the established Caco-2 cell model can be used to study the intestinal absorption mechanism of oral administration drugs.