兔胰腺干/祖细胞的分离培养和鉴定  

Isolation,Culture and Identification of Rabbit Pancreatic Stem/Progenitor Cells

作　　者：张慧茹[1] 冯若鹏[1] 刘雨潇[1] 庞全海[1] 窦忠英[1] 

机构地区：[1]国家干细胞研究中心陕西分中心西北农林科技大学动物科技学院

出　　处：《农业生物技术学报》2005年第6期768-771,i0011,共5页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No:2002AA216161);国家自然基金项目(No:302000137);教育部重点项目(No:03160)资助

摘　　要：采取1～9日龄的实验仔兔胰腺,1mg/mLⅣ型胶原酶消化,低糖DMEM培养。细胞铺满后进行传代20代以上冻存。对2～3代的细胞进行免疫组化染色,诱导向β细胞分化并进行高糖刺激胰岛素分泌。结果表明,兔的胰腺干/祖细胞具有较强的增殖能力;免疫组化染色表现为PDX-1、CK19、nestin和胰高血糖素阳性;分化诱导2d后,双硫腙染色(DTZ)20%阳性,6d后迅速增加到80%;高糖刺激诱导细胞具有胰岛素释放能力。通过免疫组化染色和诱导细胞胰岛素分泌实验证明分离的是兔胰腺干/祖细胞。首次报道兔胰腺干/祖细胞分离培养方法。The pancreas obtained from 1-9 days rabbits were digested with 1 mg/mL IV collagenase, and cultivated in low-glucose DMEM. The cells were subcultured when they grew into confluence. The cells were identified as pancreatic stem/progenitor cells with immunohistochemical staining and insulin secretion to high-glucose response after induced into β-cells. Results showed that the rabbit cells which were passaged 20 times possessed a strong proliferation ability, and the immuno histochemical positive to PDX-1, CK19, nestin and glucagon. After induced for 2 days, the positive rate of zinc-chelating dithizone （DTZ） was 20 percent, and rapidly increased to 80 percent 6 days later. The ability of insulin secretion to high-glucose respond has be performed. This test has a powerful result that these cells were proved to be pancreatic stem/progenitor cells by immunohistochemical staining and insulin release. It was the first time to report that isolation and culture of rabbit pancreatic stem/progenitor cells.

关 键 词：胰腺干细胞 兔 胰岛素 分离 鉴定 

分 类 号：S858.28[农业科学—临床兽医学] Q462[农业科学—兽医学]