A 3-D cell culture system to study epithelia functions using microcarriers

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• 作者：Petra H. Jakob ; Jessica Kehrer ; Peter Flood ; Catharina Wiegel…
• 关键词：Three ; dimensional cell culture ; Live cell imaging ; Epithelial systems ; Cytodex 3 ; EMT
• 刊名：Cytotechnology
• 出版年：2016
• 出版时间：October 2016
• 年：2016
• 卷：68
• 期：5
• 页码：1813-1825
• 全文大小：4,675 KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Biomedicine
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-0778
• 卷排序：68

文摘

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.