Fig. 1: A vascularized mammary duct platform models native 3D tissue architectures and morphogenesis. | Nature Communications

Fig. 1: A vascularized mammary duct platform models native 3D tissue architectures and morphogenesis.

From: Uncovering mutation-specific morphogenic phenotypes and paracrine-mediated vessel dysfunction in a biomimetic vascularized mammary duct platform

Fig. 1

Tissues are at one week of co-culture unless otherwise indicated. a Organotypic microfluidic device consisting of an engineered 3D endothelial vessel (green) and mammary epithelial duct (cyan) in physiologic ECM (magenta), enabling long-term culture and paracrine signaling. Inset: 3D reconstruction of endothelial vessel (green—VE-cadherin) and epithelial duct (cyan—phalloidin). b Left: Composite stitched maximum intensity projection micrograph of tissue base to medial section of a mammary epithelial duct (cyan, phalloidin) and proximal vasculature (green, phalloidin); Alexa Fluor-labeled collagen I (magenta), DAPI (blue). Top right: Representative maximum intensity micrographs of mature endothelial vessels immunostained for VE-cadherin, ZO-1, actin, and Ki67, as indicated. Bottom right: Individual confocal slice micrographs from 3D epithelial duct midsections immunostained for GM130, Ki67, α6 integrin, E-cadherin, actin, and DAPI, as is indicated in each panel. c Quantification of golgi localization as measured by the nuclear-GM130 axis (n = 52 cells examined from three independent ducts). 0 corresponds to a nuclear-GM130 axis apically oriented toward the lumen, while 180 corresponds to a basal orientation. d Cartoon depicting morphogen gradients established by perfusing growth factors through an acellular vascular channel. Immunographs of quiescent MCF10A ducts exposed to vehicle (DMSO), FGF2 (3 nM), or TGFβ1 (5 ng/ml) gradients for three days. Inset: high magnification slice micrograph of duct terminus immunostained with phalloidin (cyan), DAPI (blue) and Ki67 (magenta). e Quantification of percentage of Ki67 positive nuclei in ducts treated with each morphogen gradient (n = 4, 3, 3 ducts examined across three independent experiments; one-way ANOVA with Bonferroni post test Vehicle vs FGF2 **p = 0.0036, FGF2 vs TGFβ1 **p = 0.002, mean ± s.e.m). All images are representative of at least three independent experiments. Source Data are provided as a Source Data file.

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