Comparison of culture media for human adipose tissue mesenchymal stromal cells as an object of research and industrial cultivation

The choice of cell culturing conditions is a limiting factor for the procedures of obtaining and expanding cells in vitro, as well as for developing drugs based on them. In this work, we analyzed the characteristics of primary human mesenchymal stromal cell (MSC) lines obtained from adipose tissue, isolated and cultured in a growth medium supplemented with AdvanceSTEM™ serum (Cytiva) and a serum-free medium CellCor™ Serum free CDM for hMSC (Xcell). The use of two types of media made it possible to obtain viable MSC cell lines from human adipose tissue. At the same time, the proliferation rate in the serum-free medium was higher, which was expressed in shortening of PDT, lag phase, time between passages, both during cultivation of freshly isolated cells and after defrosting of lines initially cultured in a medium with serum. However, the use of a standard cryopreservation protocol for cells isolated and cultured in CellCor™ medium did not allow for the effective recovery of these lines from thawing. In addition, a tendency for MSCs to form clusters was found when using this medium, indicating the need for further selection of working conditions with it. MSCs are among the most promising cells for the development of products for regenerative medicine, in particular, due to the proteins secreted by these cells. The concentrations of growth factors VEGF, IGF, HGF and angiopoietin-1 in the cell-conditioned growth medium did not differ significantly when cultured in the two media. Also, no statistically significant differences were found in the concentration and size of extracellular vesicles in the medium. Thus, the use of the serum-free CellCor™ medium allows for the elimination of xenogenic products and can reduce the time of MSC cell mass production, which is an important parameter in the development of cell products, without changing the secretory properties of these cells compared to cultivation in the AdvanceSTEM™ medium, however, further selection of conditions is required to optimize the protocols.

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