Differential sensitivity of epithelial cells to extracellular matrix in polarity establishment

PLoS One. 2014 Nov 13;9(11):e112922. doi: 10.1371/journal.pone.0112922. eCollection 2014.

Abstract

Establishment of apical-basal polarity is crucial for epithelial sheets that form a compartment in the body, which function to maintain the environment in the compartment. Effects of impaired polarization are easily observed in three-dimensional (3-D) culture systems rather than in two-dimensional (2-D) culture systems. Although the mechanisms for establishing the polarity are not completely understood, signals from the extracellular matrix (ECM) are considered to be essential for determining the basal side and eventually generating polarity in the epithelial cells. To elucidate the common features and differences in polarity establishment among various epithelial cells, we analyzed the formation of epithelial apical-basal polarity using three cell lines of different origin: MDCK II cells (dog renal tubules), EpH4 cells (mouse mammary gland), and R2/7 cells (human colon) expressing wild-type α-catenin (R2/7 α-Cate cells). These cells showed clear apical-basal polarity in 2-D cultures. In 3-D cultures, however, each cell line displayed different responses to the same ECM. In MDCK II cells, spheroids with a single lumen formed in both Matrigel and collagen gel. In R2/7 α-Cate cells, spheroids showed similar apical-basal polarity as that seen in MDCK II cells, but had multiple lumens. In EpH4 cells, the spheroids displayed an apical-basal polarity that was opposite to that seen in the other two cell types in both ECM gels, at least during the culture period. On the other hand, the three cell lines showed the same apical-basal polarity both in 2-D cultures and in 3-D cultures using the hanging drop method. The three lines also had similar cellular responses to ECM secreted by the cells themselves. Therefore, appropriate culture conditions should be carefully determined in advance when using various epithelial cells to analyze cell polarity or 3-D morphogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques / methods
  • Cell Polarity*
  • Dogs
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism*
  • Extracellular Matrix / chemistry*
  • Humans
  • Madin Darby Canine Kidney Cells
  • Mice

Grant support

SY was supported by a CREST grant for “Elucidation of a novel unified epithelial system defined by the reciprocal regulation of cell-cell adhesion and the apical cytoskeleton and its use for manipulating the epithelial barrier” from the (JST) Japan Science and Technology Agency (http://www.jst.go.jp/kisoken/crest/en/project/35/e35_09.html). There is no grant number. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.