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

MACDONAL, O.; JUAN GABRIEL CHEDIACK; CAVIEDES-VIDAL, E.

San Luis

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

Sociedad de Biología de Cuyo

Evaluation of three methods for sequential isolation of avian intestinal epithelial cells. Mac donal O, Chediack JG, Caviedes-Vidal E. Departamento de Bioquímica y Ciencias Biológicas. Área de Biología. UNSL. e-mail macdonal@unsl.edu.ar A comparative study for isolating avian intestinal epithelial cells was made using mechanical, enzymatic and chelating methods. The birds used were pigeons (Columba livia) between 1-30 days old. The cells were isolated by mechanical, enzymatic and chelating methods in two steps and were characterize by morphology, cell number, viability, light micrographs and activities of villus cell marker enzymes alkaline phosphatase (AP) and sucrase- isomaltase (SI). Intestinal epithelium mass was markedly greater with enzymatic and chelating than mechanical methods. In all methods were obtained high yields of cells and high viability in both steps. Alkaline phosphatase and sucrose-isomaltase activities were constant along villus- crypt axis in each method. All methods produce a high yield of intestinal epithelial cells with viability around 50 %. Enzymes activities were not different along the intestinal villus-crypt axis, like in chickens. Therefore, unlike mammals, avian enterocytes have a similar pattern of AP and SI expression. Supported by FONCYT 1-03101 & CyT-UNSL 22/Q151 to ECV.