Fig. 6: Cross-talk between induced lung mesenchyme and epithelium impacts mesenchymal differentiation in recombinant co-cultures.
From: Directed differentiation of mouse pluripotent stem cells into functional lung-specific mesenchyme
a RT-qPCR data showing fold change expression relative to day 0 iPSCs of lung mesenchymal markers in day 11 iLM cells before co-culture (gray), and in iLM and MEFs after co-culture in distal (dark blue) and proximal (light blue) medium. Cells were re-sorted for GFP+ (iLM) or Cherry−/EPCAM− (MEFs) after 7 days of co-culture. N = 8. b Immunofluorescence images showing expression of GFP, E-CADHERIN and Smooth muscle actin (SMA) in Nkx2-1mCherry+ cells co-cultured with iLM in distal or proximal medium for 1 week. Scale bars = 100 μm. c Immunofluorescence images showing expression of GFP, NKX2-1 and PDGFRA in Nkx2-1mCherry+ cells co-cultured with iLM in distal or proximal medium for 1 week. Scale bars = 100 μm. d RT-qPCR data showing fold change expression relative to day 0 iPSCs of MANC, ASM and VSM markers in day 11 iLM cells before co-culture (gray), and in iLM after co-culture in distal (dark blue) and proximal (light blue) medium. N = 8. e SPRING plot showing sample identity and expression of Wnt2 for single cell RNA-seq of iLM before and after distal and proximal co-culture. f Dot plot showing expression of 10 distal and 10 proximal markers in iLM before and after distal and proximal co-culture. These markers were previously found to be enriched in the distal versus proximal adult mouse lung by Wang et al., 2018. g SPRING plot showing expression of gene sets (top50 expressed genes) of adult lung mesenchymal cell types described by Zepp et al., 2021. h SPRING plot showing expression of gene sets (top50 expressed genes) of adult lung mesenchymal cell types described by Tsukui et al., 2020. All bars show mean ± sd. P values were determined by paired, two-tailed Student’s t test. Significant (p < 0.05) p values are highlighted in bold font.
