摘要
本试验旨在建立马乳腺上皮细胞的原代培养方法,为马属动物泌乳相关机制的研究提供试验材料和细胞模型。以泌乳期哈萨克母马乳腺组织为研究对象,利用Ⅰ型胶原酶和透明质酸酶分离马乳腺上皮细胞,并进行纯化和鉴定。结果显示:1)利用酶消化法培养的马乳腺上皮细胞形态呈长条状或岛屿状;冻存后复苏培养的马乳腺上皮细胞生长状况良好。2)纯化后的马乳腺上皮细胞具备正常的“S”形生长周期。3)琼脂糖凝胶电泳结果显示,培养的马乳腺上皮细胞能够表达其特异性基因细胞角蛋白18(cytokeratin 18)基因(CK 18)。4)通过免疫荧光技术,运用鼠抗cytokeratin 18作为一抗,对马乳腺上皮细胞进行鉴定,结果呈阳性。综上所述,本研究使用的方法能够成功分离、纯化培养并鉴定出马乳腺上皮细胞。
The aim of this experiment was to establish a primary culture method of equine mammary epithelial cells to provide experimental materials and cell models for the study of lactation-related mechanisms in equine animals.The experiment was conducted on mammary tissue of Kazakh mares in lactation,and equine mammary epithelial cells were isolated using typeⅠcollagenase and hyaluronidase,and purified and characterized.The results showed as follows:1)the morphology of equine mammary epithelial cells cultured by enzyme digestion method was“strip-like”or“island-like”;mammary epithelial cells cultured in resuscitation after freezing were in good growth condition.2)The purified equine mammary epithelial cells had a normal“S”shape growth cycle.3)Agarose gel electrophoresis results showed that the cultured equine mammary epithelial cells were able to express their specific gene cytokeratin 18 gene(CK 18).4)The mouse anti-cytokeratin 18 was used as primary antibody to identify the equine mammary epithelial cells by immunofluorescence technique,and the result was positive.In conclusion,the method used in this study is able to successfully isolate,purify culture and identify equine mammary epithelial cells.
作者
李晓斌
李超
蔺建薇
李倩
郭庆勇
LI Xiaobin;LI Chao;LIN Jianwei;LI Qian;GUO Qingyong(Xinjiang Key Laboratory of Horse Breeding and Exercise Physiology,Key Laboratory of Herbivores for Meat and Dairy,College of Animal Science,Xinjiang Agricultural University,Urumqi 830052,China;College of Animal Medicine,Xinjiang Agricultural University,Urumqi 830052,China)
出处
《动物营养学报》
CAS
CSCD
北大核心
2023年第12期8046-8052,共7页
CHINESE JOURNAL OF ANIMAL NUTRITION
基金
新疆农业大学博士后流动站资助(2020)
自治区重点实验室开放课题(2022D04004)。
关键词
马
乳腺上皮细胞
细胞培养
鉴定
equine
mammary epithelial cells
cell culture
identification
