The impact of surgical technique on the development of graft versus host disease in a rat small intestinal transplant model.

The small intestine and its mesentery contain a large amount of lymphoid tissue that can mediate graft-versus-host disease (GVHD) in small intestinal transplant recipients. To assess the impact of surgical technique and the retention of the recipient's small intestine on GVHD intensity, 12 adult Lewis rats received heterotopic small bowel transplants and 12 received orthotopic small bowel transplants from Brown Norway donors. Twelve Lewis to Lewis heterotopic small-bowel-transplanted animals served as the control group. All recipients were given cyclosporine (10 mg/kg/alternate days) subcutaneously. The parameters followed were: weight gain and feed intake; clinical signs of GVHD; relative spleen weight; popliteal lymph node enlargement assay; and histological evaluation of spleen, liver, skin, native intestine, and transplanted intestine. According to the clinical scoring system, heterotopically transplanted animals were found to have a more severe GVHD than the orthotopic group. There were statistically significant differences between the relative spleen weights of the heterotopic transplant group and the control group (P = 0.001, 0.004, and 0.007 on days 7, 14, and 21, respectively) and between the heterotopic and orthotopic groups at 7 days (P = 0.037). Lymph node enlargement assays were statistically different between heterotopic and orthotopic groups (P = 0.019, 0.020, and 0.007 on days 7, 14, and 21, respectively). Histological evaluation of skin biopsy specimens also demonstrated that GVHD was indeed more severe in the heterotopic transplanted group when compared with orthotopically transplanted animals. These findings confirm that retention of the native small intestine in the heterotopic intestinal transplant model significantly increases the severity of GVHD following transplantation.

Cadaver pancreas recovery technique. Impact on islet recovery and in vitro function.

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.