METHODS OF UPSCALING MESENCHYMAL STROMAL CELL PRODUCTION, COMPOSITIONS AND KIT THEREOF

摘要

The present invention discloses a method of isolation, pooling and further culturing of Mesenchymal Stem cells (MSC) for clinical application. Present invention also discloses the method of establishing Master Cell bank, followed by Working Cell Bank from which the final therapeutic composition referred to as Investigational Product/Investigational Medicinal Product comprising of allogenic bone marrow-derived MSC is formulated for clinical applications. Present disclosure also discloses a robust manufacturing process for consistent production of clinical grade Mesenchymal Stromal cells (MSCs). The process enables production of highly viable potent cells. The process steps relating to preparation of media, cell seeding, harvesting are fine tuned to achieve consistency in cell yield, superior cell viability, purity, improved cell proliferation, high cell recovery, low HLA-DR expression, reduction in culture duration. The viability and purity of cells are further improved by optimized wash process without cell loss/cell stress. The disclosure further provides a method of cyrostoring MSCs at high cell density without affecting the viability of cells. It further provides economical means to store and transport at −80° C.

主权项

1. A process for culturing of mesenchymal stromal cells, said process comprising act of:
a) allowing the mesenchymal stromal cells cultured till a first predetermined passage, to expand to a second predetermined passage in presence of culture media comprising basic fibroblast growth factor (bFGF); wherein said expansion is carried out by contacting the cells with said culture media when said cells achieve at least one pre-determined confluency; and wherein the process increases number of the cells at the end of the second predetermined passage by 2000 folds when compared to number of the cells at the first predetermined passage.