Scaffold-free culture of mesenchymal stem cell spheroids …

Adipogenic differentiation potential of MSCs following 3D culture. Following 7 days of suspension culture, MSCs were recovered from mesenspheres, plated onto TCPS and maintained under growth or adipogenic culture conditions. The adipogenic differentiation potential of recovered cells was compared to that of conventional monolayer cultures. Brightfield (a, b, e, f, i, j) and phase (c, d, g, h) images of Oil Red O-stained MSCs were obtained following differentiation. a, c, e, g, i Lipid vacuole formation was not evident in any cultures maintained under growth conditions. b, d, f, h, j Lipid vacuoles were more widespread within cultures recovered from mesenspheres following 14 days of differentiation than in monolayer cultures after 28 days of differentiation. Scale bar 200 m. k Quantitative staining for adipogenesis exhibited significant differences between monolayer cultures and cells recovered from mesenspheres. Cells recovered from mesenspheres stained more robustly for Oil Red O than monolayers at day 14 of differentiation, with plated spheroids exhibiting a greater than 20-fold increase (*p< 0.0001) and dissociated spheroids exhibiting a greater than 40-fold increase (*p<0.0001) in the amount of lipid vacuole formation compared to monolayers. Dissociated mesenspheres also underwent greater adipogenesis (+p<0.0001) than plated spheroids

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Scaffold-free culture of mesenchymal stem cell spheroids ...