CHARACTERIZATION OF PRIMARY MURINE ENTEROCYTES

DI CALUDIO, F; GRIGERA, R; MUGLIA C.I.; DOCENA, G.

Los Cocos, Córdoba

Congreso; LXI Reunión Anual de la Sociedad Argentina de Inmunología; 2013

The intestinal epithelium is a highly dynamic compartment, being the enterocytes the target for different analysis. Culturing intestinal epithelial cells is difficult, and efforts are being done to improve the survival of cells. In this work we have optimized culture conditions for primary culture of murine enterocytes. Cells obtained from different regions of the gut were cultured on collagen membranes with the presence of growth factors and oligoelements in the culture medium.Cell viability, functional phenotype and proliferation were evaluated. Results showed that cells adhere to the collagen membranes and growth with an organized distribution. Acridin orange/ethidium bromide staining gave 3 +- 2 % of orange nuclei at day 7, increasing to 7 +- 2 % of orange nuclei at day 14 by fluorescence microscopy (10 fields-200X) (P menor 0.05, N=10). Annexin V/PI staining, showed by flow cytometry a low apoptosis rate (less than 3%) during the culture. The percentage of live cells remained constant (70 ± 6) at day 7, with a sharp decrease at day 14 (20 ± 3 % of live cells) (P