Fig. 2

Differentiation of MD-iPSCs into MSCs. a Schematic protocol for differentiation of MD-iPSCs into MSCs. b Formation of embryoid bodies (EBs) from MD-iPSCs. MD1- and MD2-iPSCs formed EBs with typical morphologies. Scale bars = 500 μm. c Representative images of iPSC-derived cells at 0, 1, 2, and 3 weeks after α-MEM induction. Retardation of MSC differentiation was observed both in the MD1- and MD2-iPSC groups compared with the WT-iPSC group. Scale bars = 500 μm. d Expression of MSC markers in MD-MSCs. MD1- and MD2-MSCs showed lower expression of CD105 compared with WT-MSCs. Gating strategy for this analysis is summarized in Additional file 3 (Figure S2). α-MEM α-minimum essential medium, D day of differentiation, DMEM Dulbecco’s modified Eagle’s medium, FBS fetal bovine serum, ITS ITS liquid media supplement, MD1/2 Menkes disease patient 1/2, SB SB431542, SR Knockout Serum Replacement, wk weeks, WT wild type