Separation of pure populations of epithelial cells from rabbit distal colon Academic Article uri icon

Overview

MeSH Major

  • Cell Separation
  • Colon

abstract

  • A simple method using divalent chelators is described for the isolation of viable populations of surface and crypt cells from rabbit distal colon. Histological studies were performed to monitor colonocyte dissociation and determine contamination by nonepithelial cells. Cell viability was assessed by trypan blue exclusion assay and by 22Na uptake measurements. Electron microscopy was used to determine the integrity of the isolated cells. Alkaline phosphatase and [3H]thymidine uptake were measured to assess the purity of the different cell fractions. Combined fractions 4 and 5 contained the highest percentage of pure surface cells, while fractions 10, 11, and 12 were predominantly crypts. Alkaline phosphatase activity was 13 +/- 3-fold higher in the surface cells than in the crypt cells, while [3H]thymidine uptake was 8 +/- 4-fold higher in the crypt cells than in the surface cells. Amiloride-sensitive and -insensitive 22Na uptake was the same in the surface cells directly after isolation and after 3 h in culture. In this study we demonstrate a method for the preparation of highly enriched fractions of rabbit colon surface and crypt cells that remain viable and functional in short-term culture.

publication date

  • January 1995

Research

keywords

  • Academic Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1006/abio.1995.1018

PubMed ID

  • 7710060

Additional Document Info

start page

  • 134

end page

  • 9

volume

  • 224

number

  • 1