CAJ | 학술논문

目的探索大规模猪胰岛细胞分离纯化的方法.方法联合器官切取,胶原酶P主胰管灌注,COBE2991细胞分离机及HCA-Ficoil纯化猪胰岛细胞.通过双硫腙(DTZ)染色,倒置显微镜下计数胰岛细胞的数量和纯度,胰岛素释放试验检测胰岛细胞的分泌功能.结果消化后平均每条胰腺可平均获得(275 000±20 895)胰岛细胞当量(IEQ),纯化后平均为(230 350±26 679)IEQ,平均每克胰腺组织可获得(2710±229)IEQ,纯化后胰岛细胞平均纯度为(50.2±1.95)%.纯化后的胰岛细胞对胰岛素释放刺激反应良好,高糖(16.7 mmol/L)时胰岛素的释放量为低糖(3.3 mmol/L)时的4.74倍(P≤0.001).结论成功建立了猪胰岛细胞分离、纯化的方法,纯化的猪胰岛细胞具有良好的生物活性.
목적 탐색대규모저이도세포분리순화적방법.방법 연합기관절취,효원매P주이관관주,COBE2991세포분리궤급HCA-Ficoil순화저이도세포.통과쌍류종(DTZ)염색,도치현미경하계수이도세포적수량화순도,이도소석방시험검측이도세포적분비공능.결과 소화후평균매조이선가평균획득(275 000±20 895)이도세포당량(IEQ),순화후평균위(230 350±26 679)IEQ,평균매극이선조직가획득(2710±229)IEQ,순화후이도세포평균순도위(50.2±1.95)%.순화후적이도세포대이도소석방자격반응량호,고당(16.7 mmol/L)시이도소적석방량위저당(3.3 mmol/L)시적4.74배(P≤0.001).결론 성공건립료저이도세포분리、순화적방법,순화적저이도세포구유량호적생물활성.
Objective To establish effective method for large-scale purification of islet cells from pig pan-cress. Methods Pig pancreas tissue was digested with collagenase P followed by purification in a HCA-Fi-coil dis continuous gradient using Cobe2991 cell separator. After isolation, the islet cell yield and purity were evaluated with light microscope with DTZ staining, and the islet function assessed by insulin release as-say in vitro. Results The number of the islets coll ected from each pancreas averaged (275 000±20 895)islet equivalents (IEQ) before purification, and (230 350±26 679) IEQ after the purification with discon-tinuous gradient centrifugation. From each gram of the pancreatic tissue, (2710±229) IEQ were obtained with an average purity of (50.2±1.95) %. The purified islets responded well to high-concentration (16.7 mmol/L) glucose stimulation with a 4. 74-fold increase of insulin secretion over the basal level (3.3 mmol/L, P <0.001). Conclusion The established method can be applicable for large-scale purifi-cation of fully functional islet cells from pig pancreas.