摘要
目的探索用简化的单一密度梯度法建立快速纯化大鼠胰岛细胞的方法。方法用胶原酶P消化分离SD大鼠胰腺,采用自制的推进式离心管以及单一密度(1.090g/cm3)的HCA-Ficoll-400密度梯度液来纯化大鼠胰岛细胞。通过DTZ染色,在倒置显微镜下计数胰岛细胞的数量和纯度,用胰岛素释放试验检测胰岛细胞的功能。结果纯化后平均每条SD大鼠胰腺可获得(731±89)个胰岛细胞当量(IEQ),平均纯度为(73.2±9.4)%,平均活率为(92.8±2.4)%。纯化后的胰岛细胞对胰岛素释放刺激反应良好,高糖时胰岛素的释放量为低糖时的3.54±0.79倍(P<0.001)。结论为各种胰岛细胞的实验研究建立了快速分离纯化胰岛细胞的方法,纯化的胰岛细胞功能良好。
Objective To establish an rapid and simple discontinuous density gradient method for purification of islet cells from rat pancreas. Methods SD rat pancreas was enzymaticaUy digested by collagenase P, Islet cells were purified with mono - layers of HC - A - Ficoll 400 gradient solution（ 1. 090 g/cm3 ） in modified medical syringes by mono - density gradient centrifugation method. After isolation, the islet yield and purity were evaluated with the light microscope by DTZ staining islet function was assessed by insulin release assays in vitro. Results After the purification step by modified mono - density gradient centrifugation method, about （731 ± 89） islet equality （IEQ） per pancreas was obtained from SD rat pancreas, with an average purity of （73.2 ± 9. 4） % and activity of （92. 8 ± 2, 4） %. The purified islets responded to high concentration glucose stimulation（ 16. 7 mmol/1） with （3.54 ± 0. 79） times increase of insulin secretion over basal levels（3. 33 mmol/1）. The difference was statistically signiiicant（P 〈 0. 001 ）. Conclusion We have established a simple and rapid rat islet purification method for islet study, and the purified islet cells remain full function.
出处
《广东医学》
CAS
CSCD
北大核心
2007年第3期349-351,共3页
Guangdong Medical Journal
基金
国家自然科学基金资助项目(编号:30470829)
关键词
大鼠
胰岛细胞
纯化
Rat islet cell
Purification
Mono -gradient method
