肠道转运Caco-2细胞单层模型的建立及验证评价

目的建立Caco-2细胞单层模型用于药物转运研究。方法按照常规的细胞培养方法,将Caco-2细胞接种到Millicell小室内(接种密度1×10⁶个·mL^-1),培养21 d。定期用细胞电位仪监测跨上皮细胞电阻（TEER）,评价细胞单层的紧密性与完整性;通过荧光黄转运实验检查Caco-2细胞单层模型细胞旁路转运通透性;通过普萘洛尔转运实验验证Caco-2细胞单层模型跨细胞被动转运通透性。结果培养21 d后,TEER值达到（981±123）Ω·cm²,荧光黄和普萘洛尔的表观通透系数分别为0.33×10及16.7×10^-6cm·s^-1。结论本研究建立的Caco-2细胞单层模型紧密、完整,具有良好通透性,可用于药物转运研究。

Development and validation of Caco-2 cell monolayer model of intestinal transport

AIM To develop a Caco-2 cell monolayer model for the investigation on drug intestinal transport. METHODS Caco-2 cell was seeded in Millicell plate inserts with 1 ×10^5cells· mL^- 1 cell density and cultivated in stan- dard procedures for 21 d. The value of transepithelial electrical resistance （TEER） of the monolayer was periodically monitored with a Millicell electrode resistance system （MiUipore） for the evaluation of monolayer integrity. Lucifer yellow rejection was used to examine the permeability of paracellular transport, and propranolol transport was used to verify the permeability via passive diffusion mechanism. RESULTS After 21 d, TEER value reached （981 ± 123） 12- cm2, the apparent permeability （Pare） of lucifer yellow and propranolol was 0.33 × 10^-6 cm·s^-1 and 16.7 × 10^-6cm·s^-1,re- spectively. These results showed that Caco-2 cell monolayer model developed in the present study was compact and com- plete with good permeability. CONCLUSION Caco-2 cell monolayer model is a valuable model system which can be used for the drug intestinal transport study.