Figure 8: Generation of PDMS culture inserts that mimic villus topography, and highlighting of tuft-like structures in cultures EpCAM-silenced on synthetic villi.
From: Contractile forces at tricellular contacts modulate epithelial organization and monolayer integrity
(a) Schematic representation of the synthetic polydimethylsiloxane (PDMS) villus inserts. (b,c) Scanning electron microscopy analyses of villus PDMS culture inserts: top (b) and side (c) views are presented. Scale bars, 50 μm (b) and 100 μm (c). (d,e) Confocal microscopy analysis of E-cadherin distribution in control (Caco2 shNT, upper panels) and EpCAM-silenced (Caco2 shEpCAM, lower panels) cells that were grown on 3D villus-like micropatterned PDMS inserts for 21 days. After z-stack acquisitions, 3D rendering was generated. White arrowheads point on tuft-like structures. Scale bars, 50 μm. (f) Confocal microscopy analysis of Par3, villin, actin, E-cadherin and occludin distribution in control (Caco2 shNT) and EpCAM-silenced (Caco2 shEpCAM) cells that were grown on villous PDMS inserts for 21 days. Transversal xy views are presented. Par3, villin, actin and occludin display abnormal lateral membrane localization in the absence of EpCAM (yellow arrows). Scale bars, 50 μm.
