Abstract:The Caco-2 cell model can be used to predict nutrient absorption in various ways, and it has been widely applied to the early rapid screening process of nutritional product development. Although it usually takes 21 days to establish a conventional Caco-2 monolayer model, an accelerated Caco-2 monolayer culture system was developed in this study to shorten the experimental period and reduce costs. Caco-2 cells were cultured by adding growth factors to the standard medium, and the model integrity was evaluated by its transepithelial electrical resistance (TEER) values, permeability of hydrophilic markers, expression of key protein genes, morphological characteristics of the cell membrane, etc. The results showed that the Caco-2 model obtained by the accelerated nine-day method had a TEER value of more than 220 Ω?cm2 and an apparent partition coefficient (Papp) value of leakage marker Lucifer yellow of (0.39 ± 0.15 ) × 10-6 cm/s. The apical (AP)/basolateral (BL) alkaline phosphatase activity ratio was 5.481 ± 0.5304, showing obvious polar differentiation of the cell monolayer. A quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the mRNA expression of the key proteins (P-gp and MRP2) in the cell monolayer, and no significant difference was found between the accelerated method and the standard culture method. Using this method, a complete Caco-2 cell model can be established in only nine days, providing a viable alternative to the conventional Caco-2 monolayer model.
