Abstract: Chicken oviduct epithelial cells synthesize and secret most egg albumin can be a cell bioreactor or take part in preparation of intravital bioreactor, also are a good model for studying chicken oviductal gene expression regulation. Researchers established a chicken oviduct epithelial cell line named XYF-142, meanwhile, developed a new method, obturated interior digesting for efficiently procuring uncontaminated primary cells. The population doubling time of XYF-142 was 106.67 h. About 92.5% of 5th XYF-142 cells were diploid in chromosome analysis. The results of real-time PCR indicated that the expression level of oviductal epithelia-specific gene-SERPINB3 was higher in cells than in oviduct tissue and higher in 5th and 15th epithelia than in primary epithelia. In immunofluorescence cytochemistry detection for XYF-142, all cells expressed the marker protein-cell keratin 18. XYF-142 cells were competent to be transfected with vector DNA and transfection efficiency can reach 60%. Therefore, researchers concluded that researchers have successfully established a chicken oviduct epithelialcell line, provided a model for studying gene expression regulation and tissue-specific detection of chicken oviduct.