动物营养学报2016，Vol.28Issue(4)：1225-1232,8.DOI:10.3969/j.issn.1006-267x.2016.04.034
不同培养方法对山羊瘤胃上皮细胞生长及角蛋白18表达量的影响
Effects of Different Culture Methods on Growth and Keratin 18 Expression of Ruminal Epithelium Cells of Goats
摘要
Abstract
This research aimed to study the effects of different culture methods on growth and keratin 18 ( CK18) expression of ruminal epithelium cells of goats. The test proceeded with ruminal epithelium cells of goats aged 42 days, and the cells were cultured using the methods of enzyme digestion and tissue mass culture. The morphology of cells at the stages of primary culture and subculture was observed under the optical micro-scope, and the growth curve of the fifth generation of ruminal epithelium cells of goat was tested, meanwhile, ruminal epithelium cells were identified using cell immune fluorescence method. The results showed as follows:1) the primary cultured ruminal epithelium cells of goats obtained by digestion of 0. 25% trypsin and 0. 02%ethylenediamine tetra-acetate began adherent growth on the 2nd day. There was a significant increase in cell number on the 5th day, and the cell number reached a maximum at the 10th day. 2) The primary cultured ru-minal epithelium cells of goats obtained by tissue mass culture began ‘escape’ from the tissue mass on the 4th day. There was a significant increase in cell number on the 8th day, and the cell number reached a maximum at the 14th day. 3) The CK18 in cytoplasm of cells obtained by the 2 methods exhibited a positive reaction by im-mune fluorescence staining, and cell purity was higher in tissue mass culture method. 4) Tissue mass culture method expressed significantly higher CK18 compared with enzyme method ( P<0.05) . In conclusion, ruminal epithelium cells of goats were successfully obtained with high purity using the method of tissue mass culture.关键词
山羊/瘤胃/上皮细胞/原代培养/分离/CK18Key words
goat/rumen/epithelium cell/primary culture/isolation/CK18分类
农业科技引用本文复制引用
刘思乐,康劲翮,谭支良,王征..不同培养方法对山羊瘤胃上皮细胞生长及角蛋白18表达量的影响[J].动物营养学报,2016,28(4):1225-1232,8.基金项目
国家自然科学基金项目(31201826) （31201826）
中央驻湘科研机构技术创新发展专项(2013TF3006) （2013TF3006）
