畜牧与兽医2025，Vol.57Issue(8)：36-42,7.
猪乳腺上皮细胞的分离与鉴定
Isolation and identification of porcine mammary epithelial cells
摘要
Abstract
This study was to establish a primary culture method for porcine mammary epithelial cells(PMEC),in order to provide biomate-rials and cell models for research on mammary gland development,differentiation,lactation mechanisms,and pathogen infection in pigs.The mammary gland tissue of 9-month-old non-pregnant healthy sows was used as the research object.PEMC was isolated using a combined di-gestion method involving collagenase type Ⅰ and hyaluronidase.Morphological observation,growth curve determination,immunofluor essence assay(IFA),polymerase chain reaction(PCR),and Western blot were used to identify PMECs.The results were as follows:The mammary epithelial cells obtained from enzymatic digestion exhibited a cobblestone-like and epithelioid morphology,forming single-layer clustered islands.After cry opreservation and recovery,the cells demonstrated good growth,and the purified porcine mammary epithelial cells had a normal"S"-shaped growth cycle.After stimulating mammary epithelial cells with prolactin concentrations of 0.2 mg/L and 2 mg/L,PCR and Western blot methods were used to detect a significant increase in the expression level of β-casein(CNS2)in the cells(P＜0.05),and the increasing trend was positively correlated with the concentration of prolactin.The IFA results showed that the isolated porcine mammary epithelial cells expressed the specific marker cytokeratin 18(CK-18)and tight junction protein(Zonula Occludens 1,ZO-1),indicating that the cells formed good tight junctions.In summary,in this study,a system for isolating,culturing,and identifying porcine mammary epi-thelial cells was successfully established,laying the foundation for further research on mammary biology and related mechanisms.关键词
猪乳腺上皮细胞/分离/鉴定/角蛋白18/β-酪蛋白Key words
porcine mammary epithelial cells/isolation/characterization/cytokeratin 18/CNS2分类
农业科技引用本文复制引用
翟雨薇,刘超凡,王飞燕,刘佳薇,王卫,石晓静,袁晨,宋勤叶..猪乳腺上皮细胞的分离与鉴定[J].畜牧与兽医,2025,57(8):36-42,7.基金项目
基因项目:中央引导地方科技发展资金项目(226Z6601G) （226Z6601G）
国家自然科学基金青年科学基金项目(32202843) （32202843）
河北省省属高等学校基本科研业务费研究项目(KY2022042) （KY2022042）
