Identification of optimal conditions for the expansion of human bone marrow-derived mesenchymal stem cells using tools for live-cell culture monitoring

Based on the results of our study, we have developed recommendations regarding cell culture media composition for the expansion of human bone marrow-derived mesenchymal stem cells (MSC) for preclinical studies and potential clinical applications. ALPHA-MEM supplemented with 10% platelet lysate proved to be the most effective culture medium. Different DMEM media supplemented with fetal bovine serum turned out to be less effective: a maximum of 80% confluence was reached after 80 hours of culture, while MSC confluence in StemMACS and ALPHA-MEM media supplemented with platelet lysate kept increasing even after 100 hours of expansion. The growth rate of MSCs in RPMI-1640 medium was significantly lower than in the other culture media. When culturing MSCs in media with high glucose concentration (4.5 g/L), the percentage of cells with fat transformation after 5 days of culture was higher than in low-glucose (1.0 g/L) media such as DMEM low gl, StemMacs, ALPHAMEM. It is preferable to use MSC expansion media that do not induce spontaneous adipogenic differentiation for culturing MSCs for clinical purposes because the cells remain uncommitted and all their differentiation potential can be used in accordance with the objectives of further research and/or clinical needs. This study was supported by the local Ethics Committee and approved by the Scientific Council of the Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of Ministry of Healthcare of the Russian Federation. All the participants signed the standard informed consent form and agreed to the use of some of their biological materials for research purposes.