这里,我们提出了一个病例,需要一个补充的“老派”胰岛净化,以安全的门内输注。从脑死亡的26岁男性供体(体重指数:21.9)中获取胰腺后,连续密度梯度离心后分离出24.6ml胰岛组织。胰岛产量为504,000胰岛当量(IEQ),分布在以下三个部分中:0.6ml颗粒中的64,161IEQ,182,058IEQ在10毫升,和258,010IEQ在14毫升与95%,20%,纯度10%,分别。经过23小时的培养,我们应用了补充胰岛纯化,基于单位重力沉降过程中组织亚组分的分离,一种60多年前开发的技术(“老派”)。这种方法使总颗粒体积减少到11.6毫升,同时保留374,940个IEQ,存活率超过90%。在三个输液袋中制备最终的胰岛产品,在2.6毫升颗粒中含有130,926IEQ,在4毫升颗粒中的108,079IEQ,和135,935IEQ在5毫升颗粒中含有65%,40%,纯度30%,分别,并加入普通肝素(70单位/kg体重)。从所有三个袋子中输注胰岛后,门静脉压力从7mmHg增加到16mmHg。输注后48小时继续使用肝素进行抗血栓预防,目标激活部分凝血活酶时间50-60s,然后分次皮下注射肝素2周。移植后第75天评估的β-细胞移植物功能良好,根据Igls的标准,完全消除严重的低血糖发作,胰岛素需求减少50%。在目标葡萄糖范围(70-180mg/dl)内花费的时间从42%提高到98%,HbA1c从8.7%下降到6.7%。补充“老派”胰岛纯化允许安全和成功地利用坚固和高质量的胰岛制剂,否则将被丢弃。
Here, we present a
case that required a supplemental \"old school\" islet purification for a safe intraportal infusion. Following pancreas procurement from a brain-dead 26-year-old male donor (body mass index: 21.9), 24.6 ml of islet tissue was isolated after continuous density gradient centrifugation. The islet yield was 504,000 islet equivalent (IEQ), distributed among the following three fractions: 64,161 IEQ in 0.6 ml of pellet, 182,058 IEQ in 10 ml, and 258,010 IEQ in 14 ml with 95%, 20%, and 10% purity, respectively. After a 23-h culture, we applied supplemental islet purification, based on the separation of tissue subfractions during unit gravity sedimentation, a technique developed over 60 years ago (\"old school\"). This method enabled the reduction of the total pellet volume to 11.6 ml, while retaining 374,940 IEQ with a viability of over 90%. The final islet product was prepared in three infusion bags, containing 130,926 IEQ in 2.6 ml of pellet, 108,079 IEQ in 4 ml of pellet, and 135,935 IEQ in 5 ml of pellet with 65%, 40%, and 30% purity, respectively, and with the addition of unfractionated heparin (70 units/kg body weight). Upon the islet infusion from all three bags, portal pressure increased from 7 to 16 mmHg. Antithrombotic prophylaxis with heparin was continued for 48 h after the infusion, with target activated partial thromboplastin time 50-60 s, followed by fractionated heparin subcutaneous injections for 2 weeks. β-Cell graft function assessed on day 75 post-transplantation was good, according to Igls criteria, with complete elimination of severe hypoglycemic episodes and 50% reduction in insulin requirements. Time spent within the target glucose range (70-180 mg/dl) improved from 42% to 98% and HbA1c declined from 8.7% to 6.7%. Supplemental \"old school\" islet purification allowed for the safe and successful utilization of a robust and high-quality islet preparation, which otherwise would have been discarded.