Figure 4.

Induction of MSX1⁺PDGFRA^low limb mesenchyme-like cells from human pluripotent stem cells

(A) Schematic illustration of the strategy for stepwise induction of MSX1⁺ cells from human pluripotent stem cells (hPSCs).

(B) FACS analysis confirmed the induction of MSX1⁺PDGFRA⁺ LPM-like and MSX1⁺PDGFRA^low LM-like cells from 3D-cultured MSX1^P2A-tdTomato hPSCs with FGF8+10 and mock treatments.

(C) qRT-PCR analysis confirmed the stronger induction of LM markers in the FGF8+10 treated cells. LPM makers: PDGFRA, COL1A2, COL3A1, and TSHZ2. LM markers: LHX2, HOXC10, HMMR, and MKI67. Error bars represent data from three independent experiments with triplicates. Statistics: independent-sample t test using SPSS version 22.0. ^∗∗p < 0.01 and ^∗∗∗p < 0.001.

(D) Principal component (PC) analysis of bulk RNA-seq data from hPSC-derived MSX1⁺PDGFRA^low and MSX1⁺PDGFRA⁺ cells. FGF8+10-treated cells were compared with mock control.

(E) Heatmaps of marker gene expression in FGF8+10 treated cells compared with mock control. The range of transcriptional expression is illustrated by a color change as depicted on the extreme right of the figure (dark red correlates to high expression, whereas light blue correlates to low expression).