Primary Cell Culture of Turkey Intestinal Epithelial Cells

Abstract

Primary cell culture has been widely used in various types of studies and proven useful for the isolation and identification of avian pathogens. Difficulties in growing intestinal epithelial cells in vitro have limited their use for such studies. In the present study, a co-culture system was developed for the primary culture of intestinal epithelial cells. A monolayer obtained from 14-to- 16-day-old turkey embryo intestinal fibroblasts was used as a feeder layer. Feeder layers from turkey embryo fibroblasts and from a continuous cell line (mouse 3T3 fibroblasts) were also employed but were not as successful. The intestinal epithelial cells were isolated by dissociation from the intestinal tracts of 1-day-old turkey poults and grown on the feeder layers. Growth and maintenance media were supplemented with various components, including fetal calf serum, chicken serum, hormones, and other growth factors. The epithelial cells grown on feeder layers from the intestinal fibroblasts allowed the intestinal epithelial cells to be maintained in vitro for periods of 7 to 10 days. This technique may prove useful for various applications, including isolation of enteropathogens, and for basic studies of the intestinal tract concerning such subjects as physiology, immunology, and toxicology.