Isolation, characterization, and EGFP expression in the buffalo (Bubalus bubalis) mammary gland epithelial cell line

This study was aimed to establish a buffalo mammary epithelial cells (BuMECs) line and maintain it for long-term by subculturing. BuMECs isolated from lactating buffalo mammary glands were cultured on a collagen matrix gel. BuMECs expressed significant amounts of the epithelial cell specific marker cytokeratin 18 as determined by immunohistochemistry. The BuMECs displayed monolayer, cobble-stone morphology, and formed lumen-, dome-, and duct-like structures. Furthermore, they were capable of synthesizing CSN2, BLG, ACACA, and BTN1A1, showed viability after thawing and expressed milk protein genes. The enhanced green fluorescent protein gene was transferred successfully into the BuMECs using lipofection method and the transfected cells could be maintained for long-term in culture by subculturing.