Development and characterization of a cow mammary organoid model for in vitro milk production

The primary function of the mammary gland is to produce milk for newborns. However, current in vitro models to study bovine mammary epithelium and lactation have notable limitations, particularly the loss of natural three-dimensional (3D) architecture. Mammary organoids represent an innovative solution, offering a physiologically relevant 3D model for in vitro milk production. In this context, we are developing and characterizing organoids derived from mammary epithelial stem cells cultured in extracellular matrix protein gels, which self-organize into 3D structures. A first line of research focuses on characterizing organoids under proliferative conditions (MammoCult vs Growth factor medium) using different approaches, including confocal microscopy, flow cytometry, and gene expression analysis. In parallel, a second line of research relies on single-cell transcriptomic analysis (scRNA-seq) to compare cellular and molecular profiles of organoids cultured in proliferative versus differentiative conditions (MammoCult vs progesterone/estrogen/bovine pituitary extract medium). Additionally, secretome analyses will be conducted to assess the composition and functionality of secreted factors, particularly those linked to milk production. Together, these approaches aim to establish a reproducible and functional bovine mammary organoid model for in vitro milk production.