Figure 1 |. Surface ectoderm and CNCC co-induction leads to hair-bearing skin generation.

a, b

a, b, Overview of (a) study objectives and (b) skin organoid (SkO) protocol. c, Brightfield images of WA25 aggregates on days 12 and 30 in optimized culture. d, Immunostaining for KRT5+KRT15+ basal and KRT15+ peridermal layers at day-55. e, f, Representative HF-induction images (e) in brightfield of days 6585 WA25 SkOs and (f) max-intensity confocal image (endogenous DSP-GFP) of days 6595 DSP-GFP SkO. Dashed-box: magnified-HF; dashed-line: HF; dashed-circles: developing hair germs; asterisks: dermal papilla. g, h, Violin plots showing (g) frequencies of HF-formation in WA25 (average 87.4%, min=68.8%, max=100%, n=212 organoids), DSP-GFP (average 87.2%, min=66.7%, max=100%, n=212 organoids), and WA01 (71%, n=130 organoids) cultures, and (h) average number of HFs formed in WA25 (average 64 HFs/organoid, min=9, max=285, n=80 organoids) and DSP-GFP (average 48 HFs/organoid, min=7, max=128, n=80 organoids) cultures between days 75147. i-k, Immunostained day-75 WA25 SkO with hair placodes. Antibodies highlight epidermal (KRT5+KRT15+CD49f+) and periderm (KRT15+) layers, dermis (PDGFR+P75+), and DC cells (SOX2+PDGFR+P75+). Dashed-boxes: magnified-regions. l, Wholemount of day-85 WA25 SkO with head-tail structures. KRT5 highlights epidermis and HF outer root sheath. SOX2 marks DC, DP, Merkel cells and melanocytes. Dashed-box: area shown to the right. Abbr: frontonasal prominence (FNP); cranial neural crest cells (CNCCs); dermal papilla (DP); matrix (Mtx); periderm (PD); hair root (HR); dermal condensate (DC). Scale: 500 m (c), 250 m (l; left), 100 m (e; first three panels, f; third-panel, i-k; upper-panels), 50 m (d, f; second-panel, i-k; lower-panels, l; right), 25 m (e; last-panel, f; first/last-panels). See Statistics and Reproducibility for plot and experimental information.

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Hair-bearing human skin generated entirely from pluripotent stem cells

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