Cell line/type | BM-MSCs (Bone marrow-derived mesenchymal stromal cells) |
---|---|
Species | Human |
Animal free | Yes |
Product | DMEM-PL1 Laitinen et al. 2016 |
Laitinen, A., Oja, S., Kilpinen, L., Kaartinen, T., Möller, J., Laitinen, S., ... & Nystedt, J. (2016). A robust and reproducible animal serum-free culture method for clinical-grade bone marrow-derived mesenchymal stromal cells. Cytotechnology, 68(4), 891-906. In this study, a clinically-compliant and reproducible animal serum-free culture protocol for bone marrow-(BM-) MSCs was established based on an optimized platelet-derived supplement. Our study compared two different platelet-derived supplements, platelet lysate PL1 versus PL2, produced by two different methods and lysed with different amounts of freeze-thaw cycles. Our study also explored the effect of a low oxygen concentration on BM-MSCs. FBS-supplemented BM-MSC culture served as control. Growth kinetics, differentiation and immunomodulatory potential, morphology, karyotype and immunophenotype was analysed. Growth kinetics in long-term culture was also studied. Based on the initial results, we chose to further process develop the PL1-supplemented culture protocol at 20 % oxygen. The results from 11 individual BM-MSC batches expanded in the chosen condition were consistent, yielding 6.60 x 10(9) +/- 4.74 x 10(9) cells from only 20 ml of bone marrow. The cells suppressed T-cell proliferation, displayed normal karyotype and typical MSC differentiation potential and phenotype. The BM-MSCs were, however, consistently HLA-DR positive when cultured in platelet lysate (7.5-66.1 %). A robust and reproducible clinically-compliant culture method for BM-MSCs based on platelet lysate is presented, which enables high quantities of HLA-DR positive MSCs at a low passage number and suitable for clinical use. The BM-MSC basal medium consisted of low glucose DMEM 100 U/ml penicillin, 100 µg/ml streptomycin and 40 IU/ml heparin supplemented with 10 % PL1, as specified in the Materials and Methods, under the heading; Culture of MSCs. PL1 is specified in Table 1 Characteristics of the PL1 and PL2 platelet lysate supplements and their use in MSC growth media, and Schallmoser et al. 2007. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4960139/ |
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Source | Literature - own formulation |
Chemically defined > No | No |
Contains phenol red > Yes | Yes |
Antibiotics free > No | No |