Differentiation of bmMSCs into IPCs
(A) Cell clusters were observed at various stages (5–30 days) during the differentiation process. The morphology of cells was observed in a light microscope (top panel) and a fluorescence microscope (bottom panel). (B) qPCR was used to detect mRNA expression levels of Ins1, Ins2, Pdx1, Isl-1, Glut2, Pax6, glucagon, and MafA. The level of mRNA in cells infected with controlled shRNA (shControl) plus GFP was defined as 1. MIN6 cells were used as a positive control. Data are presented as means ± SD from three independent experiments. (C) Immunofluorescence analysis for insulin, Glut2, Pdx1, and glucagon in differentiated bmMSCs infected with shRNA targeting COUP (shCOUP)-TFI plus MafA, which was visualized using confocal microscopy on day 30. DAPI was used for nuclear staining. Scale bars represent 50 μm.