不同日龄鸡的原代肝细胞分离与鉴定  

Isolation and identification of primary hepatocytes from chickens of different ages

作　　者：周静静 郭晓艳 杜瑞雪 曹亚蒙 胡薛英[1] 程国富[1] 张万坡[1] 刘晓丽[1] 谷长勤[1] ZHOU Jing-jing;GUO Xiao-yan;DU Rui-xue;CAO Ya-meng;HU Xue-ying;CHENG Guo-fu;ZHANG Wan-po;LIU Xiao-li;GU Chang-qin(College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学动物医学院,湖北武汉430070

出　　处：《中国兽医科学》2022年第10期1328-1333,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(32030107)

摘　　要：选取不同日龄肉鸡,通过剪碎法和半原位灌流法分别分离1、15、30日龄肉鸡和成年肉鸡原代肝细胞并进行培养。采用油红O染色检测脂滴存在情况,PAS染色检测糖原分泌情况,免疫组织化学法和间接免疫荧光染色法检测肝细胞特异标记物CK18。结果显示,1日龄的鸡原代肝细胞内脂滴过多,15日龄的鸡原代肝细胞状态欠佳,30日龄和成年的鸡原代肝细胞生长状态和形态良好;原代肝细胞内存在大量的脂滴和糖原,免疫组织化学法和间接免疫荧光染色显示分离的细胞为CK18阳性细胞,即为肝细胞。结果表明,30日龄和成年肉鸡采用半原位灌流法分离得到的细胞纯度高、数量多且状态良好,且分离的肝细胞具备分泌糖原和储存脂质的功能,适合进行体外试验研究。Broilers of different ages were selected in the experiment,and primary hepatocytes of 1-day-old,15-day-old,30-day-old broilers and adult broilers were isolated and cultured by shearing and semi-in situ perfusion,respectively.Lipid droplets were detected by oil red O staining,glycogen secretion was detected by PAS staining,hepatocyte specific marker CK18 was detected by immunohistochemistry and indirect immunofluorescence staining.The results showed that there were too many lipid droplets in primary hepatocytes of 1-day-old chickens,poor state of primary hepatocytes of 15-day-old chickens,good growth and morphology of primary hepatocytes of 30-day-old and adult chickens.And a large number of lipid droplets and glycogen in primary hepatocytes.Immunohistochemistry and indirect immunofluorescence staining showed that the isolated cells were CK18 positive cells,namely hepatocytes.In conclusion,the cells isolated from 30-day-old and adult broilers by semi-in situ perfusion have high purity,large number and good condition,and have the function of secreting glycogen and storing lipids,so they are suitable for research in vitro.

关 键 词：鸡 原代肝细胞 细胞分离 细胞鉴定 

分 类 号：S831[农业科学—畜牧学]