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Fig. 5 | Stem Cell Research & Therapy

Fig. 5

From: Native extracellular matrix preserves mesenchymal stem cell “stemness” and differentiation potential under serum-free culture conditions

Fig. 5

MSC self-renewal and retention of differentiation capacity after culture in SFM on TCP, BM-ECM, and CELLstart™. Early passage (P1) cultures of BM-MSCs were cultured for 7 days on TCP, BM-ECM, or CELLstart™ in SFM. At harvest, the cells were detached, counted, and re-seeded onto plastic plates for CFU assay (CFU-F, CFU-AD, and CFU-OB). Cells, before and after expansion, were plated into six-well plates in α-MEM containing 15 % FBS. After 14 days in culture, CFU-F colonies were stained with crystal violet and counted. Beginning on day 14, cultures for assay of CFU-AD were changed to adipogenic media and cultured for an additional 10 days, and the colonies counted after staining with Oil Red O. Similarly, cultures for assay of CFU-OB were changed to osteogenic media and cultured for an additional 25 days, and the colonies counted after Von Kossa staining. a The appearance and frequency of CFU-F, CFU-AD, and CFU-OB assayed at the indicated seeding density before (initial) and after 7 days of expansion on TCP, CELLstart™, and BM-ECM. b Over the course of 7 days in culture, MSC replication was represented by the fold changes in the number of colonies after expansion on the various substrates. The mean ± standard deviation was calculated on the basis of three independent experiments. *P < 0.05, versus CELLstart™; and Ɨ P < 0.05, versus TCP and initial. α-MEM alpha-minimum essential medium, BM-ECM bone marrow-derived extracellular matrix, BM-MSC bone marrow-derived mesenchymal stem cell, CFU colony-forming unit, CFU-AD colony-forming unit-adipocyte, CFU-F colony-forming unit-fibroblast, CFU-OB colony-forming unit-osteoblast, FBS fetal bovine serum, MSC mesenchymal stem cell, P1 passage 1, SFM serum-free media, TCP tissue culture plastic

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