AJP - Gastrointestinal and Liver Physiology, Vol 242, Issue 2 147-G155, Copyright © 1982 by American Physiological Society
Characterization of isolated epithelial cells from rat small intestine
F. Hartmann, R. Owen and D. M. Bissell
Intestinal mucosal cells from the rat have been isolated by a new technique
involving intravascular perfusion of an intestinal segment with
collagenase. Detached cells were flushed from the intestinal lumen with a
second perfusion circuit containing an oxygenated buffered solution with 1%
bovine serum. Sequential collection of cells at intervals during the period
of perfusion revealed that villus-tip cells are recovered first (after 15
min of collagenase perfusion), followed by midvillus (after 25 min) and
lower villus cells (after 35 min). The isolated cells were judged intact
and viable by the criteria of trypan blue dye exclusion, ultrastructural
appearance, and metabolic activity. They were characterized as villus-tip,
midvillus, and lower villus-crypt cells by their alkaline phosphatase and
sucrase activity, glycoprotein formation, and [3H]thymidine incorporation.
Microsomal monooxygenase activity was four to five times greater in
villus-tip than in lower villus cells, whereas heme oxygenase exhibited a
reverse gradient. The isolated cells synthesized heme and bilirubin under
cell culture conditions.
