Fibroblasts expressing BMP-2 increase epithelial thickness, restore cell polarity, increase planar cell divisions, and ordered differentiation to human.

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Fibroblasts expressing BMP-2 increase epithelial thickness, restore cell polarity, increase planar cell divisions, and ordered differentiation to human keratinocytes in OCs. Fibroblasts expressing BMP-2 increase epithelial thickness, restore cell polarity, increase planar cell divisions, and ordered differentiation to human keratinocytes in OCs. (A, C, E, G, H) OCs populated with control (F_GFP) or BMP-2–transduced (F_BMP2) fibroblasts were analyzed after hematoxylin and eosin staining (A) or immunostaining with Ki67 (C), ΔN-p63 (E), LAMA5 (G), and K10 (H) to reveal improved basal cell polarity, increased expression of Ki67, ΔN-p63, and LAMA5 and better suprabasal K10 expression in the presence of F_BMP-2 compared with F_GFP controls. Images are representative of three independent experiments. (B, D, F) Quantitation of epithelial thickness (B), number of Ki67+ (D), and ΔN-p63+ (F) cells in OCs populated with control (F_GFP) or BMP-2–transduced fibroblasts (F_BMP2). Error bars represent mean ± SD; statistical analysis performed using unpaired t test; n.s. not significant; *P < 0.05; ***P < 0.00. All quantitative data are derived from three independent experiments. (I, J, L, M) Immunostaining of OCs for pH 3 (green) and γ-tubulin (red) to determine keratinocyte cell division orientation in the presence of BMP-2–transduced (I, J: F_GFP, n = 28 mitoses) and control fibroblasts (L, M: F_BMP2, n = 27 mitoses). (K, N) Radial bar charts illustrating angle of keratinocyte mitoses obtained with F_BMP2 (K) and F_GFP control (N) demonstrating increased planar divisions in the latter. Lizhe Zhuang et al. LSA 2018;1:e201700009 © 2018 Zhuang et al.