Fig. 5
Long-term propagation and characterization of BN-MiKCs. a Immunofluorescence analysis of stem cell markers (KRT15, KRT14, and ITGA6) in BN-MiKCs at passage 12 and 20. Scale bars = 200 μm. b mRNA expression of keratinocyte stem cell-specific (Col17A1, GJB3, GJB2, ITGA6, KRT14, and KRT15) and HFSC-specific (ANGPTL2 and CD200) markers in pKCs, M-UCs, and BN-MiKCs at passage 4, 8, 12, 16, and 20. c Colony-forming assay of BN-MiKCs at passage 8, 12, 16, and 20. BN-iKCs were seeded at a density of 0.25 × 104 cells per well in a 6-well plate with a 3 T3-J2 feeder layer and cultured for 1 week. The plates were stained with crystal violet. d Expression of keratinocyte markers (ITGA6, KRT14, and Involucrin) in mature keratinocytes differentiated from BN-MiKCs at passage 12 and 20. Scale bars = 200 μm. Nuclei were counterstained with DAPI