The study was carried out to establish in vitro a cow mammary epithelial cell (MEC) line, able to respond to lactogenic hormones and rat tail collagen gel. Different from others’ way of harvesting MEC from digested tissue filtrate, tissue plants culture was used uniquely. Cells grew out from plants which were plated on collagen-coated plastics, at confluency, exhibited the characteristic cobblestone morphology of epithelial cells. All cells stained strongly for cytokeratin 18 but not with vimentin. Detectable quantities of α-casein and β-casein were present in the differentiation media. This study noted the establishment of a functional cow MEC line.