title: A 3-D cell culture system to study epithelia functions using microcarriers.
abstract: In vitro cell culture models used to study epithelia and epithelial diseases would benefit from the recognition that organs and tissues function in a three-dimensional (3D) environment. This context is necessary for the development of cultures that more realistically resemble in vivo tissues/organs. Our aim was to establish and characterize biologically meaningful 3D models of epithelium. We engineered 3D epithelia cultures using a kidney epithelia cell line (MDCK) and spherical polymer scaffolds. These kidney epithelia were characterized by live microscopy, immunohistochemistry and transmission electron microscopy. Strikingly, the epithelial cells displayed increased physiological relevance; they were extensively polarized and developed a more differentiated phenotype. Using such a growth system allows for direct transmission and fluorescence imaging with few restrictions using wide-field, confocal and Light Sheet Fluorescence Microscopy. We also assessed the wider relevance of this 3D culturing technique with several epithelial cell lines. Finally, we established that these 3D micro-tissues can be used for infection as well as biochemical assays and to study important cellular processes such as epithelial mesenchymal transmission. This new biomimetic model could provide a broadly applicable 3D culture system to study epithelia and epithelia related disorders.
authors: European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
authors: Petra H
authors: PH
authors: Jakob
authors: Wellcome Trust Centre for Molecular Parasitology, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK.
authors: Jessica
authors: J
authors: Kehrer
authors: School of Biology and Environmental Science, UCD Science Centre West, University College Dublin, Room 141, Science Center West, Belfield, Dublin 4, Ireland.
authors: Peter
authors: P
authors: Flood
authors: European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
authors: Catharina
authors: C
authors: Wiegel
authors: European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
authors: Uta
authors: U
authors: Haselmann
authors: Wellcome Trust Centre for Molecular Parasitology, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK.
authors: Markus
authors: M
authors: Meissner
authors: Goethe University Frankfurt, Frankfurt am Main, Germany.
authors: Ernst H K
authors: EH
authors: Stelzer
authors: School of Biology and Environmental Science, UCD Science Centre West, University College Dublin, Room 141, Science Center West, Belfield, Dublin 4, Ireland. emmanuel.reynaud@ucd.ie.
authors: Emmanuel G
authors: EG
authors: Reynaud
keywords: Cytodex 3; EMT; Epithelial systems; Live cell imaging; Three-dimensional cell culture