Improved Islet Yields From Macaca Nemestrina and Marginal Human Pancreata After Two-Layer Method Preservation and Endogenous Trypsin Inhibition

We tested whether two-layer method (TLM) pancreas preservation and trypsin inhibition (Pefabloc) during processing allows longer preservation while retaining or improving viable islet recovery. Non-marginal primate (Macaca nemestrina) and marginal human (ischemic or preservation-injured) pancreata were processed with a research-oriented pan technique (Seattle method). Organs were processed upon arrival (± Pefabloc), or after TLM or University of Wisconsin solution (UW) preservation (+ Pefabloc). Islet yield, viability, and function were assessed. Pefabloc increased M. nemestrina islet yields from 9696 ± 1749 IE/g to 15 822 ± 1332 IE/g (p \u3c 0.01). Two-layer method preservation (\u3c 6 h) further increased yields, to 23 769 ± 2773 IE/g (vs. + Pefabloc; p \u3c 0.01). Similarly, Pefabloc increased marginal human islet yields from 2473 ± 472 IE/g to 4723 ± 1006 IE/g (p \u3c 0.04). This increase was maintained after lengthy TLM preservation (\u3e 30 h; 4801 ± 1066 IE/g). We also tested the applicability of TLM preservation (23.5 ± 3.2 h) to the processing of marginal human pancreata by the Edmonton/Immune Tolerance Network clinical protocol. Islet yield and function approached published results of pancreata processed 4.8 ± 0.8 h after organ recovery (p = 0.06). Pefabloc, and TLM vs. UW preservation, prolonged the tolerable interval between organ recovery and islet isolation. Islet yield, viability, and functionality improved from both marginal and nonmarginal pancreata

Improved Islet Yields From Macaca Nemestrina and Marginal Human Pancreata After Two-Layer Method Preservation and Endogenous Trypsin Inhibition

We tested whether two-layer method (TLM) pancreas preservation and trypsin inhibition (Pefabloc) during processing allows longer preservation while retaining or improving viable islet recovery. Non-marginal primate (Macaca nemestrina) and marginal human (ischemic or preservation-injured) pancreata were processed with a research-oriented pan technique (Seattle method). Organs were processed upon arrival (± Pefabloc), or after TLM or University of Wisconsin solution (UW) preservation (+ Pefabloc). Islet yield, viability, and function were assessed. Pefabloc increased M. nemestrina islet yields from 9696 ± 1749 IE/g to 15 822 ± 1332 IE/g (p \u3c 0.01). Two-layer method preservation (\u3c 6 h) further increased yields, to 23 769 ± 2773 IE/g (vs. + Pefabloc; p \u3c 0.01). Similarly, Pefabloc increased marginal human islet yields from 2473 ± 472 IE/g to 4723 ± 1006 IE/g (p \u3c 0.04). This increase was maintained after lengthy TLM preservation (\u3e 30 h; 4801 ± 1066 IE/g). We also tested the applicability of TLM preservation (23.5 ± 3.2 h) to the processing of marginal human pancreata by the Edmonton/Immune Tolerance Network clinical protocol. Islet yield and function approached published results of pancreata processed 4.8 ± 0.8 h after organ recovery (p = 0.06). Pefabloc, and TLM vs. UW preservation, prolonged the tolerable interval between organ recovery and islet isolation. Islet yield, viability, and functionality improved from both marginal and nonmarginal pancreata

Autologous adipose-derived stem cell sheets enhance the strength of intestinal anastomosis

Objective: Adipose-derived stem cells (ASCs) are capable of multiple differentiation pathways, imparting immunomodulatory effects, and secreting factors that are important for wound healing. These characteristics can be exploited to decrease the incidence of anastomotic leakage. Methods: In order to delay local wound healing at the anastomotic site, we induced ischemia in a portion of porcine small intestine by ligating vessels. Then, we injected mitomycin C into the serosa of the small intestine above the ligated vessels. Anastomotic sites were created by 2 cm incisions made in the opposite mesenteric area. ASCs were isolated from the porcine subcutaneous fat tissues and expanded under culture conditions. ASCs were trypsinized and seeded on temperature-responsive dishes and cultured to form confluent sheets. Three ASC sheets were transplanted onto the serous membrane after suturing. The extent of anastomotic wound healing was evaluated by bursting pressure, hydroxyproline content, and mRNA expression of collagen-1 alpha1 and collagen-3 alpha1. Results: We found that transplantation of ASC sheets increased anastomotic site bursting pressure. Additionally, transplantation of ASC sheets increased the hydroxyproline content of the anastomoses. Furthermore, transplantation of ASC sheets increased mRNA expression of collagen-1 alpha1 and collagen-3 alpha1. Conclusions: Our findings showed that transplantation of autologous ASC sheets enhanced collagen synthesis and anastomotic strength. Further studies are necessary to identify substances that, in combination with ASC sheets, might enhance collagen synthesis and healing in sites of anastomosis

Pedunculated early colorectal cancer with nodal metastasis: a case report

Abstract Background Pedunculated polyps are more likely to be amenable to complete resection than non-pedunculated early colorectal cancers and rarely require additional surgery. We encountered a patient with a pedunculated early colorectal cancer that consisted of poorly differentiated adenocarcinoma with lymphatic invasion. We performed an additional bowel resection and found nodal metastasis. Case presentation A 43-year-old woman underwent colonoscopy after a positive fecal occult blood test. The colonoscopist found a 20-mm pedunculated polyp in the descending colon and performed endoscopic resection. Histopathologic examination revealed non-solid type poorly differentiated adenocarcinoma. The lesion invaded the submucosa (3500 μm from the muscularis mucosa) and demonstrated lymphatic invasion. In spite of the early stage of this cancer, the patient was considered at high risk for nodal metastasis. She was referred to our institution, where she underwent bowel resection. Although there was no residual cancer after her endoscopic resection, a metastatic lesion was found in one regional lymph node. The patient is undergoing postoperative adjuvant chemotherapy, and there has been no evidence of recurrence 3 months after the second surgery. Conclusions Additional bowel resection is indicated for patients with pedunculated polyps and multiple risk factors for nodal metastasis, such as poorly differentiated adenocarcinoma and lymphatic invasion. We encountered just such a patient who did have a nodal metastasis; herein, we report her case history with a review of the literature