摘要
为体外研究牦牛肠道粘膜免疫及肠道微生物与肠粘膜之间的互作关系提供细胞,本试验采用组织块培养法培养牦牛小肠上皮细胞(intestinal epithelial cell,IEC),差异消化法进行纯化,免疫细胞化学法进行鉴定,并用细胞计数法测定其生长曲线。组织块培养法结合差异消化法可分离纯化出牦牛小肠上皮细胞,免疫细胞化学法检测显示该细胞波形蛋白染色为阴性,角蛋白染色为阳性,阳性率达到90%;细胞生长曲线显示细胞接种1d后开始贴壁生长,3-6d进入对数生长期,从第7d开始细胞增殖基本处于停滞期。采用组织块培养法成功获得了牦牛小肠上皮细胞,证明该方法可为研究牦牛肠道免疫和肠道微生物与肠粘膜之间的互作提供牦牛小肠上皮细胞。
To provide cells for interaction between the in vitro study of yak intestinal mucosal immunity, intestinal microflora and intestinal mucosa in this experiment, intestinal epithelial cells of yak were cultured by explant culture method. The cells were purified by differential digestion, and identified by immunocytochemistry.The growth curves were determined by cell counting method. The results showed that the method of explant culture combined with differential digestion can purify intestinal epithelial cells of yak, immunohistochemistry showed that the cells were negative for vimentin staining and cytokeratin staining was positive, the positive rate reached 90%; The cell growth curve showed that the cells began to adhere to the wall after inoculation with 1 d, and 3-6 d entered the logarithmic growth phase. The cell proliferation began to stagnate at the beginning of 7 d. Yak small intestinal epithelial cells were successfully obtained by tissue mass culture, it is proved that this method can provide the yak intestinal epithelial cells for the study of yak intestinal immunity and the interaction between intestinal microorganism and intestinal mucosa.
作者
王菲
战立聪
吴庆侠
董海龙
朱洪云
WANG fe;ZHAN Licong;WU Qingxia;DONG Hailong;ZHU Hongytm(Key Laboratory of clinical veterinary medicine,college of Animal Science,Tibet Agriculture and Animal Husbandry Universit)
出处
《高原农业》
2018年第2期152-157,共6页
Journal of Plateau Agriculture
基金
2016年大学生创新性计划
西藏自治区临床兽医学高校重点实验室建设资助项目
关键词
牦牛
小肠上皮细胞
组织块培养
纯化
鉴定
yak
intestinal epithelial cell
explant culture
purification
identification
