Fig. 2.

Principal processes and technical challenges for producing cell-cultured milk using animal cell culture-based technology. The production of cell-cultured milk, found on the cultivation of mammary epithelial cells (ECs), is through a series of sequential processes as follows: (1) isolation of mammary ECs from parenchymal tissues or milk, (2) cultivation of isolated mammary ECs for the establishment of a cell line, (3) evaluation of cellular characteristics, (4) optimization of cell culture media, (5) establishment of three-dimensional cell culture system using extracellular matrix, (6) upscale of cell cultivation using stirred tank bioreactor and microcarrier, (7) down-stream processing of cell culture media, (8) production of cell-cultured milk components, and (9) processing of cell-cultured milk components. The major technical challenges for the production of cell-cultured milk are resourcing the cell line (1–3), optimizing the cell culture media (4), establishing the cell culture system (5–6), and separating milk components (7–8). Comprehensive and detailed technical challenges for cell-cultured milk production are discussed in this review