Toshihisa Kimura

Improvement of islet culture with sericin.

Islet transplantation is a promising treatment for diabetes. Serum is a necessary supplement in islet cultures, but it has various disadvantages including the risk of contamination by several pathogens. Results of this study suggest that sericin is a useful alternative supplement. Sericin accelerated the proliferation of the rat insulinoma cell line RIN-5F and improved the serum-free culture of rat islets.

Intraductal mucinous tumors occurring simultaneously in the liver and pancreas

A case of simultaneous intraductal mucinous tumors of the liver and pancreas in a 67-year-old man is described. Abdominal ultrasonography and computed tomography (CT) revealed the presence of cystic lesions with intraluminal septae both in the caudate lobe of the liver and in the uncinate process of the pancreas; these cystic lesions communicated with the hepatic duct and pancreatic duct, respectively. Mucin retention was observed in the cysts, and cholestasis was induced by mucin secretion into the common bile duct. The lesions were resected by left hepatic lobectomy with caudate lobectomy, and segmental pancreatectomy. Both lesions were multilocular cystic tumors with no papillary projections or focal mass effect in their walls. Histologically, both cystic lesions were a mixture of hyperplasia and adenoma lined by low papillary columnar epithelium. There were no cellular or histological features to suggest malignant change. The fibrous intratumor interstitium lacked any mesenchymal or ovarian-like stroma. The hepatic lesion was considered to be of a similar nature to intraductal papillary mucinous tumor (IPMT) of the pancreas. However, the two lesions occurred simultaneously in the liver and pancreas. This case is of interest in regard to the diagnosis and management of mucinous hepatopancreatobiliary lesions.

Rat islet culture in serum-free medium containing silk protein sericin

BACKGROUND The development of islet cultures is desirable for successful clinical islet transplantation. Fetal bovine serum (FBS) has been used as a supplement in islet culture medium, but it may be an unsuitable supplement due recent animal health problems. We have evaluated the use of the silk protein, sericin, derived from Bombyx mori as a replacement for FBS in islet culture medium. METHODS Twenty rat islets were cultured in medium containing either sericin or FBS, or no supplement, for 14 days, during which time viable islets were counted in order to evaluate islet survival. Insulin secretion was measured in vitro by static incubation on days 3 and 7. In vivo function of cultured islets was tested by syngeneic transplantation. The islets were evaluated histologically and immunohistochemically after culture and transplantation. RESULTS Ninety-five percent of islets were viable after culture for 14 days in culture medium supplemented with either FBS or sericin, while no islets survived beyond 7 days in culture without supplement. No significant differences in stimulated insulin secretion were noted between two groups of islets grown on supplemented media. Following transplantation, islets cultured in FBS or sericin rapidly reversed hyperglycemia and maintained normal glycemic control. Histologically, islets cultured with sericin displayed a well-preserved structure and strong insulin staining before and after transplantation. CONCLUSION Serum-free medium containing sericin appears to be useful for islet culture.

EFFECTS OF MICRO-ENCAPSULATION ON MORPHOLOGY AND ENDOCRINE FUNCTION OF CRYOPRESERVED NEONATAL PORCINE ISLET-LIKE CELL CLUSTERS

Background. For the success of clinical islets transplantation, the development of a long-term storage method is necessary.However, the structure of digested islets is scanty for culture and cryopreservation. In this study, the effect of micro-encapsulation to cryopreserved porcine islet-like cell clusters (ICCs) was investigated. Methods. The ICCs prepared from neonatal pigs by collagenase digestion and culture technique were cryopreserved and micro-encapsulated in 5% agarose membranes. After cryopreservation, ICC cultured without encapsulation (group A) and cultured with encapsulation (group B) were assessed by comparison with no cryopreserved ICC (control) both in vitro by static incubation test and in vivo in a xenotransplantation study. Results. Micro-encapsulation was able to maintain the fine morphology and the number of ICCs of group B after 7 days of culture. There were not significant differences in insulin secretion of group B and control on day 1 and 7 of culture (1 day:11±0.99, 7 days: 5.30±1.08 &mgr;U/ICC/hr NS versus control). On day 7 of culture, the retrieval rate of group B (105.2±9.8%) is obviously higher compared with group A (63.0±6.3%). In the xenotransplatation model, the ICCs of group B showed long survival time (7.9±0.4 weeks) and good transplantation effect. Conclusion. Our study suggests that micro-encapsulation is one of the useful method for cryopreserved ICC to maintain the fine morphology and effectively recover the endocrine function.

Revascularization and function of pancreatic islet isografts in diabetic rats following transplantation.

In pancreatic islet transplantation, revascularization is crucial for the grafts survival and function. In this study, the endothelium of isolated islets and revascularization and function of islet isografts in diabetic rat were investigated. Islets were isolated from Lewis rats by collagenase digestion method and were examined using immunohistochemistry (CD31 stain) on days 0, 1, 3, and 7 after isolation. The number of CD31-positive cells in these isolated islets was counted (mean ± SD%). Isografts (freshly isolated islets: group A, and islets cultured for 7 days: group B) transplanted in the renal subcapsule of streptozotocin-induced diabetic Lewis rats were examined using immunohistochemistry (CD31 stain) on days 3, 5, and 7 after transplantation. Intravenous glucose tolerance tests (IVGTT) were performed on days 3 and 7 after transplantation. The number of CD31-positive cells in the isolated islets on days 0, 1, 3, and 7 after isolation were: 17.3 ± 4.1%, 8.2 ± 0.7%, 2.1 ± 0.8%, and 0.8 ± 0.5%, respectively (p < 0.05). On day 5 after transplantation, CD31-positive cells were not detected in group A and B grafts, but were detected in both groups in periphery of the islets. On day 7, CD31-positive microvessels were present throughout the entire graft. IVGTT values in groups A and B on days 3 and 7 after transplantation did not show significant differences. In renal subcapsular isografts in diabetic rats, revascularization into islet grafts occurs from the surrounding host tissue 5 days after transplantation, but has no influence on the response to glucose during this period.

Mesenteric Castleman's disease: report of a case.

Abstract.A 77-year-old woman was admitted with intermittent abdominal dull pain. Hypochromic anemia, hypergammaglobulinemia, and elevated C-reactive protein were found in this case. Ultrasonography, computed tomography, magnetic resonance imaging, and angiography indicated either mesenteric leiomyoma or leiomyosarcoma, but no definitive preoperative diagnosis could be established. A surgical resection of the tumor revealed a mesenteric Castlemans tumor with small daughter lymphoid tumors of plasma cell type.

Effects of bafilomycin A1, a vacuolar type H+ ATPase inhibitor, on the thermosensitivity of a human pancreatic cancer cell line

Purpose: It has been known that the thermosensitivity of tumour cells can be increased by lowering intra-cellular pH (pHi) by inhibiting pHi control mechanisms. The pHi is partially controlled by transport of H+ from cytoplasm into endocytic and secretary systems in the cells mediated by vacuolar type H+ATPase and also by transport of H+ through plasma membrane. Methods: This study investigated the effects the bafilomycine A1, an inhibitor of the vacuolar type H+ATPase and the EIPA, an inhibitor of the Na+/H+ exchanger in plasma membrane, on thermosensitivity of AsPC-1 cells, a human pancreatic cancer cell line. It also investigated the effects of combination of bafilomycine A1 and EIPA. Results: The treatment of cancer cells with bafilomycine A1 or EIPA individually slightly lowered pHi of the cells in vitro and increased the thermosensitivity of the cells. Conclusion: The combination of these two drugs significantly lowered pHi and increased thermosensitivity of cancer cells in vitro and enhanced the heat-induced the growth delay of AsPC-1 tumours grown s.c in the legs of BALB/cA nude mice.

Improving function and survival of porcine islet xenografts using microencapsulation and culture preconditioning.

Introduction and Aims Porcine pancreatic islets have been difficult to preserve because isolated porcine islets tend to disaggregate to single cells and lose function under culture conditions. In the current study, the influence of agarose microencapsulation on the maintenance of the number and function of islets in culture preservations and the effect of culture preconditioning of microencapsulated porcine islets on xenogenic transplantation were investigated. Methodology Porcine islets were isolated and then microencapsulated in 5% agarose membrane. The percentage of naked and microencapsulated islets remaining in the culture preservations was assessed. The effect of microencapsulation and culture on secretory function was investigated in vitro. The survival of overnight-cultured and 7-days-cultured microencapsulated islets in xenogenic transplantations was examined. Results A good percentage of microencapsulated islets remained in the culture preservations. They could maintain good secretory functions in vitro after 7 days of culture. In addition, we observed a significant prolongation of mean islet survival by culture preconditioning. Conclusions The present findings suggest that microencapsulation is one of the useful preserving methods for maintenance of the number and function of cultured isolated porcine islets. Moreover, culture preconditioning is effective for improving islet survival and might be a good option leading to clinical success.

Preservation of pancreatic islets in rodent models.

Islet-cell transplantation has some advantages over vascularized pancreas transplantation, but data from clinical islet cells transplantation have shown that some serious problems need to be overcome. One of them is the storage of islet cells. We investigated methods of preserving islet cells using culture-preservation and cryopreservation. Cryopreservation is thought to be effective for long-term preservation of large quantities of islet cells, because they can be cryopreserved without loss of their physiologic activity using a relatively rapid cooling rate of 25 degrees C/min. Moreover, in the allogenic transplantation models of cryopreservation of dissociated islet cells, there was a significant prolongation of survival time. These results suggest that cryopreservation of islet cells involves not only variable preservation methods but may also lead to a modification of graft immunogenicity.

Pancreaticoduodenal artery aneurysm involving papilla of vater successfully treated with endovascular approach despite intraoperative rupture.

An 81-year-old woman presented with dizziness and nasal bleeding. Gastrointestinal fiberscopy (GIF) showed a pulsatile aneurysm in the duodenum, and that the orifice of the papilla of Vater was involved. Three-dimensional computed tomography imaging showed an unruptured aneurysm in the pancreatic duodenal arcade. The patient underwent an emergent endovascular embolization of the donor arteries using coils and gelatin sponge particles. She was discharged without any complications. This case was extremely rare because of the anatomic location of the unruptured pancreaticoduodenal artery aneurysm and the fact that it involved the papilla of Vater, was detected with GIF, and was successfully treated endovascularly.