Study on Primary Culture and Identification of Cecum Epithelial Cells from Chicken Embryo

To establish an in vitro model for studying the injury mechanism of E.tenella,in vitro culture techniques of cecum epithelial cells,the host cells of E.tenella in chicken,were studied.Primary cecum epithelial cells from chicken embryo were isolated by the digestion of trypsin,collagenaseⅠ,thermolysin,thermolysin/collagenaseⅠ and dispaseⅠ/collagenaseⅪ,respectively.The best method for separating chicken embryo cecum epithelial cells was screened out by determining the cell viability,proportion of cell aggregates,cell total yield and cell aggregates yield.Thereafter cell purification,culture and identification were performed.The results showed that the best methods for isolating and purifying cecum epithelial cells were digesting the cecum from chicken embryo with thermolysin,then removing single cells by low-speed centrifuge and fibroblast cells by differential velocity adherent.The isolated and purified cells obtained using the above method stayed in logarithmic growth phase from the 3rd to 5th day and from the 10th to 11th day after planting,respectively,and could survive for more than 14 days.The cultured cells were identified as cecum epithelial cells by morphological observation,alkaline phosphatase staining and scanning electron microscope observation.The proportions for epithelial cells at the 4th,7th and 11th day were 81.67%,84.33% and 72.00%,respectively,which implied that the primary cecum epithelial cells with high purity could be obtained from chicken embryo by this method.