一种改良的大鼠胰岛细胞原代培养方法  

Improved method of primary culture of pancreatic islet β cell

作　　者：邓秀玲[1] 陈璐璐[1] 袁莉[1] 周慜[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院内分泌科,湖北武汉430022

出　　处：《中国现代医学杂志》2006年第7期1025-1027,1036,共4页China Journal of Modern Medicine

基　　金：教育部留学回国人员科研启动基金(2002247);湖北省自然科学研究基金(2002AB136);武汉市科技晨光计划研究基金(99100209)

摘　　要：目的探讨一种简单易行的大鼠胰岛细胞培养方法,并寻求进行体外干预实验的最适条件。方法选用3、4周龄SD大鼠胰腺,用0.5mg/mL的V型胶原酶逐级消化分离,80目尼龙筛网滤过消化产物后,加入含有11.1mmol/L葡萄糖的RPMI1640培养液内,进行体外培养。DTZ染色、利用抗胰岛素抗体进行免疫细胞染色。应用放射免疫技术对培养液上清液中的胰岛素进行检测和分析。结果胰岛细胞可以在体外培养2周以上。培养初期,细胞形态及分泌功能较稳定。DTZ染色β细胞胞浆着色,为均一的猩红色。Insulin抗体染色β细胞胞浆着色,经DAB显色为棕黄色。随培养时间延长,胰岛素分泌逐渐下降,培养7d内,胰岛素分泌较稳定。结论改良后的大鼠胰岛细胞培养方法简单可靠,细胞培养至3～7d时,适合用于体外实验研究。[Objective] To explore a convenient and reliable method to isolate and culture rat islet cell, and to seek for its best situation for experiments in vitro. [Methods] The pancreas of SD rats aged 3～4 w were digested gradually by type V collagenase. The digested tissue was filtered through a nylon mesh, collected and incubated in RPMI 1640 containing 11.i mmol/L glucose. DTZ stain and anti-insulin immunocytochemistry stain were used to identify β cens. Insulin secretion was detected by radioimmunoassay technique. [Results] Pancreatic cens could be cultured for more than2 w in vitro. DTZ stain and anti-insulin immunocytochemistry stain both showed the color in cell cytoplasm. Insulin secretion reduced with the days of culturing, a relatively stable secretion was displayed in the first week. [Conclusion] Our method of islet cell culture is simple and reliable. The ceils incubated during 3～7 day are available for experimental study.

关 键 词：胰岛细胞培养 Β细胞 胰岛素 

分 类 号：Q813.11[生物学—生物工程]