The establishment and culture of bovine omasal epithelial cells (BOECs) in vitro is a valuable tool for the study of the physiological function, nutrient absorption, and transport mechanisms of the omasum in dairy cows. This paper proposes a method for the culture of primary BOECs. Trypsin digestion was used to subculture the BOECs, which were passaged for 20 generations in vitro, and showed typical epithelial-like characteristics and a cobblestone morphology. The primary BOECs had a fast growth phase (between days 4 and 5) and were validated by their slight β-galactosidase and visible cytokerat in 18 expression. In addition, RT-PCR results demonstrated that the monocarboxylate transporter 1 (MCT1), Na+/H+exchanger 1 (NHE1), and Na+/H+ exchanger 3 (NHE3) were expressed in the isolated primary BOECs. In conclusion, this primary BOEC isolation and culture model is a promising method for the study of nutrient absorption and regulation, as well as the immune regulation of epithelial cell transport in vitro.
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