ESTABLISHMENT AND VALIDATION OF A DRUG IN VITRO ABSORPTION MODEL OF MDCK-MDR1 CELLS

Abstract

Objective To establish and validate a drug in vitro absorption model of MDCK-MDR1 cells for research on the mechanism of oral drug absorption and transport. Methods MDCK-MDR1 cells with different concentrations (group L with 1.0×108/L, group M with 2.5×108/L, and group H with 5.0×108/L) were inoculated onto a 24-well Transwell plate and cultured for 1-7 d. Optical density (OD) was measured to plot the growth curve of MDCK-MDR1 cells, and cell morphology was observed at different time points of culture. Transepithelial electrical resistance (TEER) was measured at different time points to determine the optimal cell inoculation concentration and culture time for the formation of a monolayer structure. Lucifer yellow transfer assay was used to validate the monolayer structure formed at different concentrations and time points. Results The groups L, M, and H showed the formation of the monolayer structure on days 5, 3, and 1, respectively, after inoculation, and OD reached the peak value on days 5, 4, and 3, respectively, For group L, TEER reached 300 Ω·cm2 on day 5 of inoculation and tended to be stable on days 5-7. Therefore, 1.0×108/L was selected as the optimal cell inoculation concentration, with an optimal culture time of 5 d. Lucifer yellow transfer assay showed that the monolayer structure had a Lucifer yellow apparent permeability coefficient of 4.27×10-7 cm/s, which was lower than 5.0×10-7 cm/s determined by the permeability test. Conclusion The MDCK-MDR1 cell model established in this study has validated monolayer structural integrity and permeability and can be used as an in vitro model to simulate the absorption and transport mechanisms of oral drugs.

Keywords

• mdck-mdr1 cells|electric impedance|cell membrane permeability|models,biological|biological transport|intestinal absorption|in vitro techniques