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Fig. 3 | Biological Procedures Online

Fig. 3

From: A simple and reliable protocol for long-term culture of murine bone marrow stromal (mesenchymal) stem cells that retained their in vitro and in vivo stemness in long-term culture

Fig. 3

Increased osteogenic differentiation capacity in BMSCs-FS as compared with ST2 cells. a Osteoblast differentiation of BMSCs-FS (p25) as compared with ST2 cell line. Cells were induced with osteogenic cocktail and osteogenesis was measured by quantitative alkaline phosphatase activity (ALP) and (b) Alizarin red staining. ALP and Alizarin red measurements were normalized to the cell number. ALP and Alizarin red staining images are shown. c qPCR analysis of the osteogenic markers mRNA expression after 12 days of induction in BMSCs-FS compared to ST2 cell line. Each target gene was normalized to reference genes and presented as fold change over control induced. Date were represented as fold change over control non-induced. d Up-regulated osteogenic genes in BMSCs-FS versus ST2 cells after 6 days of osteogenic induction. Gene expression was measured by qPCR using osteogenic RT2 profiler array as described in M&M. Values are mean ± SD of three independent experiments, (*p < 0.05, **p < 0.005, compared to ST2 cells at the same time points). Columns of the same group with different letters at each time point indicate significant differences according to Duncan’s multiple range tests at p < 0.05

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