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. 2016 Feb 18;6:21747. doi: 10.1038/srep21747

Figure 3. Effects of 42 °C culture on iPS cells in co-culture with other cells.

Figure 3

(a) Human iPS cells cultured on MEF were cultured at 42 °C for 1 or 2 days. Upper panels are representative of phase-contrast images. Bars, 100 μm. Middle panels are representative images of Oct4 (green) and Hoechst (blue) staining. Bars, 200 μm. Lower, montage images of 81 fields (9 × 9) of Oct4 staining images (original magnification of each field is ×20). Multiple images from the same sample were acquired using the same microscope settings. (b) The Oct4(+) and Oct4(-) cell number in 81 fields at each time point was calculated and shown in the graph (n = 3). (c) Co-culture experiments between iPS cells and iPS cell-derived cardiac cells. Two days after starting co-culture of cell aggregates of iPS cells with iPS cell-derived cells, cells were cultured at 37 °C or 42 °C for 2 days in AK03 or 10% FBS DMEM. Upper panels are representative of phase-contrast images. Bars, 100 μm. Lower panels are representative of immunofluorescent images (Oct4; green, cTnT; red). Nuclei were stained with Hoechst (blue). Bars, 200 μm. (d) The Oct4(+) cell number in 36 fields at each time point was calculated and shown in the graph (n = 4). *p < 0.05 vs. day 0. **p < 0.01 vs. day 0.