Co-culture of two MDCK strains with distinct junctional protein expression

a model for intercellular junction rearrangement and cell sorting

C B Collares-Buzato, M A Jepson, G T McEwan, B H Hirst, N L Simmons

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Distinct epithelial MDCK cell strains displaying extremes in transepithelial electrical resistance (paracellular permeability) have been established in co-culture and the subsequent cellular behaviour and formation of junctional complexes investigated. After high-density seeding, MDCK strain I and II cells in co-culture are initially randomly distributed but subsequently sort themselves out in a time-dependent manner to form separate homotypic aggregates. The final pattern of cell arrangement of homotypic aggregates depends on the relative seeding proportion of each cell type. Immunostaining of established marker proteins for junctional complexes has revealed that MDCK I and II cells differ in the degree of expression of the zonula-adherens-associated protein, E-cadherin, their cytoskeletal architecture and the junctional distribution of a desmosomal protein, and by showing subtle differences in tight junction staining for the zona-occludens-associated proteins, ZO-1 and occludin. The distinct pattern of junctional protein expression is maintained when the two MDCK strains are co-cultured; however, morphologically atypical intercellular junctions between heterotypic cells at the boundary of homotypic cell aggregates have been observed. It has been suggested that cell sorting, a phenomenon yet to be completely understood, is involved in important morphogenetic processes. We propose that co-culture of strains of the well-characterised MDCK cell line may be a novel but well-defined cell system for studying epithelial cell rearrangement and sorting in intact epithelial sheets.
Original languageEnglish
Pages (from-to)267-76
Number of pages10
JournalCell and Tissue Research
Volume291
Issue number2
Publication statusPublished - Feb 1998

Fingerprint

Intercellular Junctions
Coculture Techniques
Proteins
Madin Darby Canine Kidney Cells
Zonula Occludens Proteins
Epithelial Cells
Occludin
Adherens Junctions
Tight Junctions
Cadherins
Electric Impedance
Permeability
Staining and Labeling
Cell Line

Keywords

  • Animals
  • Cadherins
  • Cell Adhesion
  • Cell Adhesion Molecules
  • Cell Aggregation
  • Cell Membrane Permeability
  • Coculture Techniques
  • Dogs
  • Epithelial Cells
  • Intercellular Junctions
  • Kidney
  • Membrane Potentials
  • Membrane Proteins
  • Microscopy, Electron
  • Microscopy, Fluorescence
  • Morphogenesis
  • Occludin
  • Phosphoproteins
  • Phosphorylation
  • Protein Processing, Post-Translational
  • Protein-Tyrosine Kinases
  • Zonula Occludens-1 Protein

Cite this

Co-culture of two MDCK strains with distinct junctional protein expression : a model for intercellular junction rearrangement and cell sorting. / Collares-Buzato, C B; Jepson, M A; McEwan, G T; Hirst, B H; Simmons, N L.

In: Cell and Tissue Research, Vol. 291, No. 2, 02.1998, p. 267-76.

Research output: Contribution to journalArticle

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T2 - a model for intercellular junction rearrangement and cell sorting

AU - Collares-Buzato, C B

AU - Jepson, M A

AU - McEwan, G T

AU - Hirst, B H

AU - Simmons, N L

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N2 - Distinct epithelial MDCK cell strains displaying extremes in transepithelial electrical resistance (paracellular permeability) have been established in co-culture and the subsequent cellular behaviour and formation of junctional complexes investigated. After high-density seeding, MDCK strain I and II cells in co-culture are initially randomly distributed but subsequently sort themselves out in a time-dependent manner to form separate homotypic aggregates. The final pattern of cell arrangement of homotypic aggregates depends on the relative seeding proportion of each cell type. Immunostaining of established marker proteins for junctional complexes has revealed that MDCK I and II cells differ in the degree of expression of the zonula-adherens-associated protein, E-cadherin, their cytoskeletal architecture and the junctional distribution of a desmosomal protein, and by showing subtle differences in tight junction staining for the zona-occludens-associated proteins, ZO-1 and occludin. The distinct pattern of junctional protein expression is maintained when the two MDCK strains are co-cultured; however, morphologically atypical intercellular junctions between heterotypic cells at the boundary of homotypic cell aggregates have been observed. It has been suggested that cell sorting, a phenomenon yet to be completely understood, is involved in important morphogenetic processes. We propose that co-culture of strains of the well-characterised MDCK cell line may be a novel but well-defined cell system for studying epithelial cell rearrangement and sorting in intact epithelial sheets.

AB - Distinct epithelial MDCK cell strains displaying extremes in transepithelial electrical resistance (paracellular permeability) have been established in co-culture and the subsequent cellular behaviour and formation of junctional complexes investigated. After high-density seeding, MDCK strain I and II cells in co-culture are initially randomly distributed but subsequently sort themselves out in a time-dependent manner to form separate homotypic aggregates. The final pattern of cell arrangement of homotypic aggregates depends on the relative seeding proportion of each cell type. Immunostaining of established marker proteins for junctional complexes has revealed that MDCK I and II cells differ in the degree of expression of the zonula-adherens-associated protein, E-cadherin, their cytoskeletal architecture and the junctional distribution of a desmosomal protein, and by showing subtle differences in tight junction staining for the zona-occludens-associated proteins, ZO-1 and occludin. The distinct pattern of junctional protein expression is maintained when the two MDCK strains are co-cultured; however, morphologically atypical intercellular junctions between heterotypic cells at the boundary of homotypic cell aggregates have been observed. It has been suggested that cell sorting, a phenomenon yet to be completely understood, is involved in important morphogenetic processes. We propose that co-culture of strains of the well-characterised MDCK cell line may be a novel but well-defined cell system for studying epithelial cell rearrangement and sorting in intact epithelial sheets.

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KW - Cadherins

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KW - Cell Aggregation

KW - Cell Membrane Permeability

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KW - Intercellular Junctions

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KW - Membrane Potentials

KW - Membrane Proteins

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KW - Microscopy, Fluorescence

KW - Morphogenesis

KW - Occludin

KW - Phosphoproteins

KW - Phosphorylation

KW - Protein Processing, Post-Translational

KW - Protein-Tyrosine Kinases

KW - Zonula Occludens-1 Protein

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EP - 276

JO - Cell and Tissue Research

JF - Cell and Tissue Research

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ER -