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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1×10~6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·cm~2,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~(-6)cm·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。
Objective To establish a Caco-2 cell monolayer model for drug delivery studies. Methods Caco-2 cells were inoculated into Millicell cells (inoculation density 1 × 10 ~ 6 · mL -1) according to the conventional cell culture method and cultured for 21 days. Periodontal epithelial cell resistance (TEER) was periodically monitored by cell potentiometry to evaluate the tightness and integrity of the cell monolayer. Cell viability of Caco-2 cell monolayer model was examined by fluorescence yellow transfer assay. Transient transport experiments validated the transcellular permeability of Caco-2 monolayers across cells. Results The TEER value reached (981 ± 123) Ω · cm ~ 2 after 21 days of culture. The apparent permeability coefficients of Fluorescent Yellow and Propranolol were 0.33 × 10 and 16.7 × 10 ~ (-6) cm · s ~ -1). Conclusion The Caco-2 cell monolayer model established in this study is compact, complete and has good permeability and can be used for drug transport research.