Figure 4.

Increased Mineralization of FOP hiPSC-Pericytes

(A) Representative Alcian blue staining of the micromass for 3 days chondrogenic differentiation is shown. Scale bar, 2 mm.

(B) Alcian blue staining of micromasses was quantified by stain extraction and absorbance reading at 595 nm.

(C) ALP staining was performed 14 days (D14) after maintaining in α-MEM with 10% FBS (Ctrl) and osteogenic medium (OM). Results are shown as representative scanned images (left) and images taken at 4× magnification (right). Scale bar, 200 μm.

(D) The mineralization was visualized by 2% alizarin red S staining on day 21 (D21) and day 28 (D28). Results are shown as representative scanned images (left) and images taken at 4× magnification (right). Scale bar, 200 μm.

(E) Alizarin red S staining on D21 and D28 was quantified by stain extraction and absorbance reading at 570 nm.

(F) Relative gene expression analysis of ALP, COL1α, and OSC on day 14 and day 28 of osteoblast differentiation. All experiments were normalized to ACTIN. Control, the average of the results from C3-3 and UE017C1; FOP, the average of the results from F2-6 and F3-8.

(G) ALP assay on day 7 of F3-8 after maintaining in OM pretreated with LDN-212854 is shown.

Data are presented as mean and SD from four times independent experiments in (E) and three times in (B) and (G).