Evaluation of three methods for sequential isolation of avian intestinal epithelial cells.

MAC DONAL, O.; CHEDIACK, J.G.; ENRIQUE CAVIEDES VIDAL

Merlo, Mendoza

Congreso; Reunión Científica Anual de la Sociedad de Biología de Cuyo; 2004

EVALUATION OF THREE METHODS FOR SEQUENTIAL ISOLATION OF AVIAN INTESTINAL EPITHELlAL CELLS. Mac Dona1 0, Chediack JG, Caviedes- Vidal E. Unidad Invest Cs Biol, Univ Nac San Luis, 5700 Sun Luis. E-mail: macdonal@unsl.edu.ar We evaluated three methods (mechanical, enzymatic and by chelating agents) for isolation of intestinal epithelial cells. We selected Feral pigeons (Columba livia) as the biological model. We used birds of 1 to 30 days old. Cells were isolated by mechanical, enzymatic and chelating methods in two steps and characterized by morphology, cell number, viability, microphotographs and the activities of alkaline phosphatase (AP), a villus cell marker and sucrase- isomaltase (SI). Intestinal epithelial mass was markedly greater with the enzymatic and chelating procedures than with the mechanical method. In all methods high yields of cells and high viability in both steps were obtained. AP and SI activities were constant along villus- crypt axis with each method. All processes produced a high number of intestinal epithelial cells with viability around 50%. As it happens in chickens, enzyme activities were not different along the intestinal villus-crypt axis, Therefore, unlike mammals, avian enterocytes have a similar pattern of AP and SI expression.