Despite the increasing demand for milk, there is a simultaneous growth in awareness regarding sustainable dairy farming and concerns about environmental issues. The concept of generating milk components without traditional dairy farming has been introduced through the utilization of bovine mammary cells. However, the establishment of a robust primary bovine mammary alveolar cells for cell-cultured milk component production remains a challenge. Hence, the aim of this study was to assess the cellular attributes and milk component productivity of primary bovine mammary cells through various stages of cell subculture. The 1 cm