畜牧兽医学报2017，Vol.48Issue(7)：1212-1220,9.DOI:10.11843/j.issn.0366-6964.2017.07.005
绒山羊附睾上皮细胞培养方法的建立及其生长特性
Establishment of Culture Method and Growth Characteristics for Epididymal Epithelial Cells from Cashmere Goats
摘要
Abstract
The aim of this study was to establish a simple and feasible in vitro culture system for high purity epididymal epithelial cells from cashmere goat.The primary cells were obtained from epididymal caput of 10-12 months cashmere goat using enzymatic dispersion and improved tissuepiece digestion method.The cells were identified by immunocytochemistry and immunofluorescence methods.Cell purity and proliferation were also investigated respectively by flow cytometry and CCK method.Ultrastructures of epididymal epithelial cells were observed under transmission electron microscope.The results showed that the cells obtained by both methods were in islandlike clonal growth and good proliferation ability,and possessed active secretory and metabolic function.The expression of epithelial special cytokeratin 18 and epididymal special glutathione peroxidase 5 (GPX5) and the result of flow cytometry showed that both methods could obtain high purity epididymal epithelial cells.The primary cells obtained by improved tissue-piece digestion method took a long culture time,compared to that the primary cells obtained by enzymatic dispersion method which took a short culture time with more cells.These results provided a good foundation for further study on the function of epididymal epithelial cells of cashmere goat.关键词
细胞培养/附睾上皮细胞/酶消化法/改良组织块消化法Key words
cell culture/epididymal epithelial cells/enzymatic dispersion method/improved tissue-piece digestion method分类
农业科技引用本文复制引用
范晓梅,张通,粟瑞兰,边小娜,张家新..绒山羊附睾上皮细胞培养方法的建立及其生长特性[J].畜牧兽医学报,2017,48(7):1212-1220,9.基金项目
国家自然科学基金(31460598) （31460598）
海南省重大科技计划(ZDKJ2016017-02) （ZDKJ2016017-02）
