Plasma Rich in Growth Factors Induces Cell Proliferation, Migration, Differentiation, and Cell Survival of Adipose-Derived Stem Cells

Figure 4

AKT mediates PRGF survival and prevention of H₂O₂ cytotoxicity. (a) Human ASCs were treated with 100 μM H₂O₂ for 24 h in the presence or absence (0%) of 2.5% PRGF or 2.5% HS, and the cell viability was analyzed by MTT assay. The significant reduction on cell viability induced by hydrogen peroxide was prevented by 2.5% of PRGF. versus 0 μM H₂O₂ and ^# versus 100 μM H₂O₂. (b) Preincubation with 10 μM of the AKT inhibitor (Calbiochem, VIII 124018) abolished the cell protective effect of PRGF of the cytotoxic effects of 100 μM H₂O₂. versus 0% PRGF. (c) FACS analysis of annexin V showed the percentage of apoptotic cells according to each treatment. ^# versus 2.5% PRGF. (d) Western blot analysis of total protein lysates of the human ASCs treated with 2.5% PRGF (+) or 0% PRGF (−) in the presence of 100 μM, preincubated or not for 30 min with 10 μM AKT inhibitor for 24 h. Activation or cleavage of the apoptotic protein PARP is confirmed when two bands visible. β-Actin was employed as a loading control. Representative blots of three different experiments are shown; .

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