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. 2021 Oct 5;9:747521. doi: 10.3389/fcell.2021.747521

FIGURE 3.

FIGURE 3

Impact of donor age on mesenchymal stromal cells (MSC) osteogenic differentiation in vitro. (A) Alkaline phosphatase (ALP), von Kossa (VK), Alizarin Red, osteopontin (OPN), and osteocalcin (OC) stainings of young MSC (30 and 45 years old) after 21 days of culture under osteogenic conditions. (B) ALP, VK, Alizarin Red, OPN, and OC stainings of old MSC (60 and 80 years old) after 21 days of culture under osteogenic conditions. DAPI was used to counterstain the cell nuclei in blue. DMEM: cells before differentiation (day 0), OSTEO: cells after 21 days of culture under osteogenic differentiation conditions. (C) ALP activity of young and old MSC after 21 days under osteogenic differentiation conditions. (D) Calcium deposition quantification of young and old MSC. (E) Osteogenic gene expression by young MSC and old MSC after 21 days under osteogenic differentiation conditions (Col I, Runx2, ALP, OPN, OC). Results were normalized to the endogenous control GAPDH and presented as fold change expression relative to undifferentiated MSC (day 0). Values are expressed as mean ± SD (two donors/group, three independent experiments/donor); *p < 0.05; Scale bars, 100 μm.