Kneteman N M, Lakey J R, Kizilisik T A, Ao Z, Warnock G L, Rajotte R V
Department of Surgery, University of Alberta, Edmonton, Canada.
Transplantation. 1994 Nov 27;58(10):1114-9.
Previous studies in rodent and canine animal models have suggested a detrimental impact on islet recovery and function when pancreas excision is preceded by in situ vascular flushing with cold preservation solutions. We studied the efficacy of islet isolation from 19 consecutive cadaver pancreases procured alternately by initial pancreatectomy before in situ flush (group 1, our standard procurement technique, n = 9) or pancreas removal following in situ vascular flushing with cold University of Wisconsin solution (group 2, n = 10). Once procured, pancreases were weighed, the main pancreatic duct was cannulated, and 150 ml of collagenase solution was injected. The pancreases were transported to the isolation laboratory and processed within 2 hr. Islets were counted and sized after dithizone staining, and the islet equivalents were calculated. Aliquots of isolated islets were cryopreserved using established techniques. Islet function of both freshly isolated and frozen-thawed islets was assessed using a glucose-stimulated perifusion system. Significantly more pancreas was harvested after University of Wisconsin flush (90.6 +/- 6.9 g for group 1 versus 66.7 +/- 4 for group 2, P < 0.05). The quantity of islets per gram of processed pancreas released during enzymatic digestion from each of the experimental groups did not differ significantly (4.5 +/- 0.6 x 10(3) islet equivalents per gram for primary pancreatectomy versus 4.0 +/- 0.4 x 10(3) University of Wisconsin flush). Similarly, following Ficoll purification, the overall yields of islets did not differ significantly. Total islet yield in the primary pancreatectomy group was 181 +/- 25 x 10(3) islet equivalents (2.7 +/- 0.3 x 10(3) IE/g) versus 217 +/- 41 x 10(3) for the University of Wisconsin flush group (2.9 +/- 0.8 x 10(3) islet equivalents/g; P not significant). No differences were observed in in vitro viability. Perifusion stimulation indexes (peak/basal insulin release) were 5.9 +/- 1.3 for group 1 and 7.1 +/- 1.5 for group 2. These results conflict with published results in animal models and indicate that large numbers of viable islets can be recovered from cadaver pancreas utilizing either procurement technique. The decreased operating time, simplicity, and safety favor the use of total pancreatectomy after limited in situ vascular flushing as the method of choice for pancreas procurement for subsequent islet isolation.
以往在啮齿动物和犬类动物模型中的研究表明，在原位用冷保存液进行血管冲洗后再进行胰腺切除，会对胰岛恢复和功能产生不利影响。我们研究了从19个连续的尸体胰腺中分离胰岛的效果，这些胰腺通过两种方式交替获取：一种是在原位冲洗前先进行胰腺切除术（第1组，我们的标准获取技术，n = 9）；另一种是在用冷的威斯康星大学溶液进行原位血管冲洗后再切除胰腺（第2组，n = 10）。获取胰腺后，称其重量，将主胰管插管，并注入150毫升胶原酶溶液。胰腺被运至分离实验室，并在2小时内进行处理。用双硫腙染色后对胰岛进行计数和测量大小，并计算胰岛当量。使用既定技术对分离出的胰岛等分试样进行冷冻保存。使用葡萄糖刺激灌流系统评估新鲜分离和冻融后胰岛的功能。威斯康星大学冲洗后收获的胰腺明显更多（第1组为90.6±6.9克，第2组为66.7±4克，P < 0.05）。每个实验组在酶消化过程中每克处理过的胰腺释放的胰岛数量没有显著差异（初次胰腺切除术组为每克4.5±0.6×10³个胰岛当量，威斯康星大学冲洗组为4.0±0.4×10³个）。同样，经过菲可分离后，胰岛的总产量没有显著差异。初次胰腺切除术组的胰岛总产量为181±25×10³个胰岛当量（2.7±0.3×10³个IE/g），而威斯康星大学冲洗组为217±41×10³个（2.9±0.8×10³个胰岛当量/g；P无显著性差异）。在体外活力方面未观察到差异。灌流刺激指数（峰值/基础胰岛素释放）第1组为5.9±1.3，第2组为7.1±1.5。这些结果与动物模型中已发表的结果相矛盾，表明使用任何一种获取技术都可以从尸体胰腺中回收大量有活力的胰岛。手术时间缩短、操作简单和安全性高，使得在有限的原位血管冲洗后进行全胰腺切除术作为后续胰岛分离的胰腺获取首选方法。
