Fig. 3
Establishment of a protocol for the differentiation of iPSC-derived Prg4-mRFP1-positive cells. a Schematic representation of the protocol used for the differentiation of iPSC-derived Prg4-mRFP1-positive cells. Embryoid bodies (EB) were cultured in suspension for the first 5 days and adhered to culture plates after that. Different differentiation factors were supplied. b Representative bright-field and fluorescence images of embryonic stem cells (ESCs) and iPSCs (#2–9) undergoing differentiation on days 5, 12, and 21. Scale bars: 200 μm (day 5) and 50 μm (days 12 and 21). c Quantification of iPSC-derived Prg4-mRFP1-positive cells on day 21. mRFP1-positive cells were counted after single cell dissociation (left figures). Their average on day 21 was 2.2% (maximum 7.0%). The mean of 13 independent experiments is shown. Scale bars: 50 μm