Evaluation of three methods for sequential isolation of epithelial cells from avian intestine

MAC DONAL, O; CHEDIACK, JG; CAVIEDES-VIDAL E

BIOCELL

Lugar: Mendoza; Año: 2008 vol. 32 p. 219 - 227

0327-9545

The isolation of viable enterocytes, villi and crypts from the small intestine of a wild bird (Columba livia) is important to perform physiological experiments in membrane transport processes with ecological relevance. A comparative study for isolating epithelial cells, villi and crypts from avian intestine was made using mechanical, enzymatic and chelating agents. The objectives were the isolation of enterocytes, villus and crypts from the small intestine of young pigeons; the evaluation of the viability of the intestinal epithelial cells isolated; and to verify the integrity of enterocytes by biochemical features. Enzymatic and mechanical methods yielded both elongated columnar and spherical cells. In the chelating method we obtained villus and crypt units. All methods produce a high yield of intestinal epithelial cells with viability around 50 %. Brush border enzymes (sucrase-isomaltase and alkaline phosphatase) activities were high and were not different along the intestinal villus-crypt axis, in despite of chickens. Even though three methods have good viabilities, the enzymatic technique give better yield in the number of cells while the chelating method provides enriched populations of morphologically distinctive villus and crypt units.