TRANSFECTION OF EXOGENOUS DNA IN CHICKEN OVIDUCT CELLS IN VITRO

Abstract

Effective production of pharmaceutical proteins using transgenic poultry may be achieved only from a construction of expression vector for chicken oviduct bioreactor. This study was focused on development of temporary expression system by using primary oviduct epithelial cells in which transfected gene expression can be studied. The present work was aimed to study three manners for preparation of primary oviduct cells, but only one was effective. Selective digestion of chicken oviduct tissue pieces in trypsin at 38° C and mechanical withdrawal of upper cell layer resulted in the isolation of purity primary culture. Experiments indicated that monolayer cultured for 2-3 d were fit to be passaged. Then after 1-2-d, the cells were transfected. Based on our result with primary culture, we did not used complicated enzyme treatment and extra equipment in order to increase the purity of epithelial cells. Liposomes «Lipofectine® 2000» were used for primary oviduct cells transfection of a plasmid designed based on the pIRES EGFP2 vector (Clontech, United States). The ratio of cells carrying GFP activity was 5-8% of the total number of cultured cells. This researches was focused on development in vitro of temporary expression system in which transfected gene expression can be tested for 4-5 d.