beta-Defensins are important components of innate immunity in mucosal tissue, a major entry site for several pathogens. These small cationic peptides possess antimicrobial activity against various microorganisms including Salmonella. Two chicken inbred lines, 6 and 15I, diverge phenotypically with respect to levels of Salmonella Enteritidis intestinal carriage and to level of gene expression of two beta-defensins, AvBD1 and AvBD2. The cellular source of these two defensins in the intestinal tissue has not previously been explored. Therefore embryonic intestinal cells were isolated from both chicken lines. Primary intestinal cell cultures expressed epithelial specific markers (villin and E-cadherin) and differentially expressed two beta-defensin genes AvBD1 and AvBD2 according to chicken line. Furthermore, S. Enteritidis interfered with AvBD2 expression only in the cells from the susceptible line 15I. Our embryonic cell culture model demonstrated that intestinal epithelium express beta-defensin antimicrobial peptides that may play a role in immunoprotection against Salmonella Enteritidis.