Fig. 4

Antioxidants preserve ADSC cell stemness and multidirectional differentiation potential during long-term in vitro expansion. After treatment with 10 μM GSH or melatonin, the ADSCs cultured for passage 3 (P3), passage 6 (P6), and passage 9 (P9) were used in the following analysis. a Osteogenesis differentiation of passaged ADSCs (Alizarin Red S staining; scale bar, 50 μm). b Adipogenesis differentiation of passaged ADSCs (Oil Red O staining; scale bar, 50 μm). c Western blot analysis for RUNX-2 in osteogenic cells. d Western blot analysis for perilipin A in adipogenic cells. e Chondrogenesis differentiation of passaged ADSCs (Alcian blue staining; scale bar, 50 μm). f Western blot analysis for SOX-9 in chondrogenic cells. g Western blot analysis for SOX-2, OCT-4, and β-actin in ADSCs. ADSCs, adipose tissue-derived stem cells; GSH, reduced glutathione