Methods of upscaling mesenchymal stromal cell production, compositions and kit thereof

摘要

This invention relates to an improved process for culturing of bone marrow derived mesenchymal stromal cells, wherein one of the major technical features of the invention lies in the protocol devised to ensure uniform distribution of bFGF in the culture medium. This, along with the specific seeding and feeding schedule described therein, ensures efficient expansion of cell by at least 2000 fold within merely two consecutive passages. The process comprises the steps of: allowing the mesenchymal stromal cells cultured till a first predetermined passage, to expand to a second predetermined passage by consecutive passages in presence of culture media comprising uniform distribution of basic fibroblast growth factor (bFGF) at a concentration of about 2ng/ml; wherein said expansion is carried out by contacting the cells with said culture media when said cells achieve at least one pre-determined confluency; wherein the process increases number of the cells at the end of the second predetermined passage by at least 2000 folds when compared to number of the cells at the first predetermined passage; and wherein the first predetermined passage ranges from passage 3 to passage 8; and the second predetermined passage ranges from passage 5 to passage 10.