Primary culture of cat intestinal epithelial cells in vitro and the cDNA library construction
会议名称：《2021中国畜牧兽医学会兽医寄生虫学分会第16次学术研讨会》
会议日期：2021年
学科分类：090601[农学-基础兽医学] 09[农学] 0906[农学-兽医学]
摘 要：Objective To lay a foundation for screening the ***-felids interaction factors,we have developed a reproducible primary culture method for cat intestinal epithelial cells(IECs).Methods The primary IECs were isolated from a new born cat s small intestine jejunum region without food ingress,and respectively in vitro cultured by tissue cultivation and combined digestion method with collagenase XI and dispase I,then purified by *** identification,the ds cDNA of cat IECs was synthesized for constructing pGADT7 homogenization three-frame plasmid,and transformed into the yeast Y187 for generating the cDNA *** Our results indicated that cultivation of primary cat IECs relays on combined digestion to form polarized and confluent monolayers within 3 days with typical features of normal epithelial *** purified cells cultured by digestion method were identified to be nature intestinal epithelial cells using immunohistochemical analysis and were able to maintain viability for at least 15 *** homogenizable ds cDNA,which is synthesized from the total RNA extracted from our cultured IECs,distributed among 0.5-2.0 kb,and generated satisfying three-frame cDNA library with the capacity of 1.2 × 10～6 and the titer of 5.2 × 10～7 pfu/*** Our results established an optimal method for the culturing and passage of cat IECs model in vitro,and laid a cDNA library foundation for the subsequent interaction factors screening by yeast two-hybrid.