Lakey J R, Warnock G L, Rajotte R V, Suarez-Alamazor M E, Ao Z, Shapiro A M, Kneteman N M
Department of Surgery, and the Surgical-Medical Research Institute, University of Alberta, Canada.
Transplantation. 1996 Apr 15;61(7):1047-53. doi: 10.1097/00007890-199604150-00010.
In an attempt to reduce the variability in the yields of human islets isolations and to identify donor factors that were potentially deleterious, we retrospectively reviewed 153 human islets isolations in our center over a 3-year period. Isolations were performed using controlled collagenase perfusion via the duct, automated dissociation, and Ficoll purification. Factors leading to successful isolations (recovery of >100,000 islet equivalents at a purity >50%) were analyzed retrospectively using univariate and multivariate analysis. Critical factors in the multiorgan cadaveric donors that were identified using univariate analysis included donor age (P<0.01), body mass index (BMI)(P<0.01), cause of death (P<0.01), and prolonged hypotensive episodes (systolic blood pressure <90 mmHg or mean arterial pressure <60 mmHg for > 15 min) requiring high vasopressors (>15 microgram/kg/min dopamine or >5 microgram/kg/min Levophed) (P>0.01). Independent analysis of 19 donor variables using multivariate logistic stepwise regression showed six factors were statistically significant. Odds ratio (OR) showed that donor age (OR 1.1, P<0.01), local procurement team (OR 10.9, P<0.01), and high BMI (OR 1.4, P<0.01) had a positive correlation with islet recovery. In contrast, hyperglycemia (all blood glucose >10 mmol/L) (OR 0.63, P<0.01), frequency and duration of cardiac arrest (OR 0.7, P<0.01), and increased duration of cold storage before islet isolation (OR 0.83, P<0.01) had negative correlation. Using these combinations of factors, the prediction of success was 85% accurate. By donor age, success was 13% for 2.5- to 18-year-old donors (n=23), 37% for 19- to 28-year-old donors (n=30), 65% for 29- to 50-year-old donors (n=70), and 83% for 51- to 65-year-old (n=29) donors. However, when vitro function was assessed by perifusion, the insulin secretory capabilities of islets isolated from the >50-year-old donor group was significantly reduced as compared with the 2.5- to 18-year-old group (P<0.02). Multiple regression analysis using postdigestion and postpurification islet recovery as outcome variables identified BMI, procurement team, pancreas weight, and collagenase digestion time factors tht can affect the recovery of human islets. Locally procured pancreases and donors with elevated minimum blood glucose levels were identified as factors that affect the insulin secretory capabilities of the isolated islets. This review of parameters suggests an improved approach to the prediction of successful islet isolation from human pancreases. Selection of suitable pancreases for processing may improve consistency in human islet isolation and thereby decrease costs.
为了减少人类胰岛分离产量的变异性，并确定可能有害的供体因素，我们回顾性分析了本中心在3年期间进行的153例人类胰岛分离病例。胰岛分离采用经导管控制胶原酶灌注、自动解离和菲可分离法。采用单因素和多因素分析对成功分离（纯度>50%时回收>100,000个胰岛当量）的因素进行回顾性分析。单因素分析确定的多器官尸体供体中的关键因素包括供体年龄（P<0.01）、体重指数（BMI）（P<0.01）、死亡原因（P<0.01）以及需要高剂量血管升压药（多巴胺>15微克/千克/分钟或去甲肾上腺素>5微克/千克/分钟）的长时间低血压发作（收缩压<90 mmHg或平均动脉压<60 mmHg超过15分钟）（P>0.01）。使用多因素逻辑逐步回归对19个供体变量进行独立分析，结果显示有6个因素具有统计学意义。优势比（OR）表明，供体年龄（OR 1.1，P<0.01）、当地采购团队（OR 10.9，P<0.01）和高BMI（OR 1.4，P<0.01）与胰岛回收率呈正相关。相比之下，高血糖（所有血糖>10 mmol/L）（OR 0.63，P<0.01）、心脏骤停的频率和持续时间（OR 0.7，P<0.01）以及胰岛分离前冷藏时间延长（OR 0.83，P<0.01）呈负相关。利用这些因素组合，成功预测的准确率为85%。按供体年龄划分，2.5至18岁供体（n=23）的成功率为13%，19至28岁供体（n=30）的成功率为37%，29至50岁供体（n=70）的成功率为65%，51至65岁供体（n=29）的成功率为83%。然而，当通过灌流评估体外功能时，与2.5至18岁组相比，从>50岁供体组分离的胰岛的胰岛素分泌能力显著降低（P<0.02）。以消化后和纯化后的胰岛回收率作为结果变量进行多元回归分析，确定了BMI、采购团队、胰腺重量和胶原酶消化时间等因素会影响人类胰岛的回收率。本地采购的胰腺和最低血糖水平升高的供体被确定为影响分离胰岛胰岛素分泌能力的因素。对这些参数的回顾提示了一种改进的方法来预测从人类胰腺成功分离胰岛。选择合适的胰腺进行处理可能会提高人类胰岛分离的一致性，从而降低成本。
