Figure 6
Osteo-chondrogenic commitment of SBA iPSC lines. (A) qRT-PCR analysis for pluripotency (OCT4), chondrogenic (SOX9, COL2A1, ACAN) and folate receptor 1 (FOLR1) genes at day 0 and day 7 of chondrogenic induction is shown as relative expression to housekeeping gene GAPDH in CTRL iPSCs and SBA-derived chondrocytes. NS, not significant. Data are plotted as average ± SEM. *P < 0.05 CTRL iPSC d7 vs SBA d7, CTRL iPSC + FA d7 vs SBA d7, SBA d7 vs SBA + FA d7 (ACAN). N = 4 independent experiments per iPSC line. (B) ACB staining of micromass cultured from CTRL iPSC and SBA-derived chondrocytes and negative CTRL (CTRL iPSC without chondrogenic induction) at day 7 of chondrogenic differentiation. Representative images of CTRL1#2 and SBA2#1 are shown. Scale bar = 1 mm. (C) Quantification of CTRL iPSC and SBA-derived chondrocytes at day 7 of chondrogenic differentiation, measured by ICN Titertek Multiskan Plus. Data are plotted as average ± SEM. **P < 0.01 CTRL iPSC vs SBA, CTRL iPSC vs SBA + FA, CTRL iPSC + FA vs SBA + FA. N = 4 independent experiments per iPSC line. (D) qRT-PCR data of CTRL iPSCs and SBA-derived osteoblasts for pluripotency (OCT4), osteogenic (RUNX2, OCN) and folate receptor 1 (FOLR1) genes at day 0 and day 21 of osteogenic differentiation is shown as relative expression to housekeeping gene GAPDH. NS, not significant. Data are plotted as average ± SEM. **P < 0.01 CTRL iPSC d21 vs SBA d21, SBA d21 vs SBA + FA d21 (OCN). N = 4 independent experiments per iPSC line. (E) ARS staining of CTRL iPSC and SBA-derived osteocytes and negative CTRL (CTRL iPSC without osteogenic induction) at day 21 of osteogenic differentiation. Representative images of CTRL1#2 and SBA2#1 are shown. Scale bar = 100 μm. (F) Quantification of CTRL iPSC and SBA-derived osteoblasts at day 21 of osteogenic differentiation, measured by ICN Titertek Multiskan Plus. Data are plotted as average ± SEM. **P < 0.01 CTRL iPSC vs SBA, CTRL iPSC + FA vs SBA, *P < 0.05 SBA vs SBA + FA. N = 4 independent experiment per iPSC line.
