Intraductal collagenase delivery into the human pancreas using syringe loading or controlled perfusion
Intraductal collagenase delivery into the human pancreas using syringe loading or controlled perfusion
复制标题
DOI：
10.1177/096368979900800309
复制
发表时间：
1999-05-01
影响因子：
3.3
通讯作者：
Rajotte, RV
中科院分区：
文献类型：
--
作者：
Lakey, JRT;Warnock, GL;Rajotte, RV
将胶原酶有效地通过导管内输送至胰腺对于随后分离活胰岛的能力至关重要。大多数临床胰岛移植中心在胰管插管后使用注射器逆行注射将酶加载到胰腺中。另一种方法是使用再循环灌注装置系统通过胰管用胶原酶溶液灌注胰腺。这提供了对灌注压力和胶原酶温度的控制。本研究报告了我们对将 Liberase(TM)-HI 输送到 14 名连续成人多器官尸体捐献者的胰腺中的评估。使用相同的方案获取和处理替代腺体，但胶原酶递送除外。使用灌注技术对第一组胰腺进行负载，其中将冷的（4摄氏度）Liberase TM -HI以80mmHg灌注5分钟，之后将压力增加至180mmHg。然后将se胶原酶溶液缓慢升温至35℃，转移至解离室并机械解离，然后使用Ficoll的不连续梯度进行纯化。第二组中的胰腺在机械分离和纯化之前使用注射器技术加载胶原酶（28-32℃）。两组供者的胰腺冷缺血、供者年龄、体重指数、最高血糖或血清淀粉酶均无显着差异。消化室中胶原酶的平均消化时间在两组之间没有差异；然而，注射器加载组中解离后剩余的未消化组织量显着更高（15.3 +/- 2.6 g vs. 4.6 +/- 2.1 g，平均值+/- SEM，p < 0.05）。灌注组胰岛消化后恢复为 471 +/- 83 x 10(3) LE，而注射器加载组为 391 +/- 57 x 10(3) LE。灌注组的纯化后回收率较高（379 +/- 45 vs. 251 +/- 28 x 10(3) IE，p < 0.05，双尾配对 t 检验）。葡萄糖灌注后两组之间的体外胰岛活力没有观察到差异，灌注组的计算刺激指数为 4.6 +/- 0.6，注射器加载组为 4.2 +/- 0.7。通过胰管的受控灌注可以有效地将酶输送到胰腺的所有区域，从而实现最大扩张，从而增加胰岛的恢复，并且对随后的体外胰岛功能没有不利影响。
Effective intraductal delivery of the enzyme collagenase into the pancreas is crucial to the subsequent ability to isolate viable islets. Most clinical islet transplant centers load the enzyme into the pancreas by retrograde injection using a syringe following cannulation of the pancreatic duct. An alternative approach is to perfuse the pancreas via the pancreatic duct with collagenase solution using a recirculating perfusion device system. This provides control over perfusion pressures and collagenase temperature. This study reports on our evaluation of the delivery of Liberase(TM)-HI into the pancreas of 14 consecutive adult multiorgan cadaveric donors. Alternate glands were procured and processed using an identical protocol with the exception of collagenase delivery. The first group of pancreases was loaded using the perfusion technique where cold (4 degrees C) Liberase(TM)-HI was perfused at 80 mmHg for 5 min after which the pressure was increased to 180 mmHg. The se collagenase solution was then slowly warmed to 35 degrees C, transferred to the dissociation chamber and mechanically dissociated, and then purified using discontinuous gradients of Ficoll. Pancreases in the second group were loaded with collagenase (28-32 degrees C) using the syringe technique before mechanical dissociation and purification. Then were no significant differences in pancreas cold ischemia, donor age, body mass index, maximum blood glucose, or serum amylase of the donors between the two groups. Mean collagenase digestion time in the digestion chamber was not different between the two groups; however, the amount of undigested tissue remaining after dissociation was significantly higher in the syringe-loaded group (15.3 +/- 2.6 g vs. 4.6 +/- 2.1 g, mean +/- SEM, p < 0.05). Postdigestion recovery of islets was 471 +/- 83 x 10(3) LE in the perfusion group compared with 391 +/- 57 x 10(3) LE for the syringe-loaded group. Postpurification recovery was higher in the perfused group (379 +/- 45 vs. 251 +/- 28 x 10(3) IE, p < 0.05, two-tailed paired t-test). No difference in in vitro islet viability was observed between the two groups following glucose perifusion with the calculated stimulation index of 4.6 +/- 0.6 for the perfusion group and 4.2 +/- 0.7 for the syringe-loaded group. Controlled perfusion via the pancreatic duct allows the effective delivery of the enzyme achieving maximal distension to all regions of the pancreas leading to an increased recovery of the islets with no detrimental effect on subsequent in vitro islet function.