畜牧兽医学报2009，Vol.40Issue(11)：1699-1704,6.
鸡胚盲肠上皮细胞原代培养与鉴定
Study on Primary Culture and Identification of Cecum Epithelial Cells from Chicken Embryo
摘要
Abstract
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, col-lagenase Ⅰ, thermolysin, thermolysin/collagenase Ⅰ and dispase Ⅰ/collagenaseⅪ, respectively. The best method for separating chicken embryo cecum epithelial cells was screened out by deter-mining 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 em-bryo with thermolysin, then removing single cells by low-speed centrifuge and fibroblast cells by differ-ential velocity adherent. The isolated and purified cells obtained using the above method stayed in loga-rithmic growth phase from the 3rd to 5th day and from the 10th to llth 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 llth day were 81.67%, 84.33% and 72.00%, respectively, which implied that the primary cecum epithelial cells with high purity could be ob-tained from chicken embryo by this method.关键词
盲肠上皮细胞/细胞分离/体外培养/鉴定/鸡胚Key words
cecum epithelial cells/ cell separation/ in vitro culture/ identification/ chicken embryo分类
农业科技引用本文复制引用
古少鹏,王敏霞,赵素芬,郑明学..鸡胚盲肠上皮细胞原代培养与鉴定[J].畜牧兽医学报,2009,40(11):1699-1704,6.基金项目
国家自然基金资(30771619) （30771619）
山西省自然基金(20041094) （20041094）
山西省攻关项目(2006031059) （2006031059）
