scholarly journals Comparison of Trypsin Inhibitors in Preservation Solution for Islet Isolation

2009 ◽  
Vol 18 (5-6) ◽  
pp. 541-548 ◽  
Author(s):  
Hirofumi Noguchi ◽  
Michiko Ueda ◽  
Shuji Hayashi ◽  
Naoya Kobayashi ◽  
Teru Okitsu ◽  
...  
Keyword(s):  
Islet Transplantation ◽  
Stimulation Index ◽  
Islet Isolation ◽  
Preservation Solution ◽  
University Of Wisconsin ◽  
Trypsin Inhibition ◽  
Clinical Islet Transplantation ◽  
Pancreas Preservation ◽  
Collagenase Inhibition

Islet transplantation has recently emerged as an effective therapy and potential cure for type 1 diabetes mellitus. Recent reports show that the two-layer method (TLM), which employs oxygenated perfluorochemical (PFC) and University of Wisconsin (UW) solution, is superior to simple cold storage in UW for pancreas preservation in islet transplantation. Moreover, we recently reported that islet yield was significantly higher in the ET-Kyoto solution with ulinastatin (MK)/PFC preservation solution compared with the UW/PFC preservation solution in the porcine model and that the advantages of MK solution are trypsin inhibition and less collagenase inhibition. In this study, we compared ulinastatin with another trypsin inhibitor, Pefabloc, in preservation solution for islet isolation. Islet yield before purification was higher in the MK/PFC group compared with the ET-Kyoto with Pefabloc (PK)/PFC group. The stimulation index was higher for the MK/PFC group than for the PK/PFC group. These data suggest that ET-Kyoto with ulinastatin was the better combination for pancreas preservation than ET-Kyoto with Pefabloc. Based on these data, we now use ET-Kyoto solution with ulinastatin for clinical islet transplantation.

scholarly journals Excellent Islet Yields after 18-h Porcine Pancreas Preservation by Ductal Injection, Pancreas Preservation with MK Solution, Bottle Purification, and Islet Purification Using Iodixanol with UW Solution and Iodixanol with MK Solution

2019 ◽  
Vol 8 (10) ◽  
pp. 1561 ◽  
Author(s):  
Kazuho Kuwae ◽  
Chika Miyagi-Shiohira ◽  
Eri Hamada ◽  
Yoshihito Tamaki ◽  
Kai Nishime ◽  
...  
Keyword(s):  
Stimulation Index ◽  
Islet Isolation ◽  
Preservation Solution ◽  
Porcine Pancreas ◽  
Porcine Islet ◽  
University Of Wisconsin ◽  
Pancreas Preservation ◽  
Purification Methods ◽  
Pancreas Weight ◽  
Wisconsin Solution

Successful islet isolation is the key to successful islet transplantation. Our group recently modified the islet isolation protocol to include pancreatic ductal injection of the preservation solution, pancreas storage in modified extracellular-type trehalose-containing Kyoto (MK) solution, and use of an iodixanol-based purification solution and bottle purification. In this study, we applied these methods to porcine islet isolation after 18-h pancreas preservation and compared two solutions with different compositions in bottle purification. Islet yield before purification was 651,661 ± 157,719 islet equivalents (IE) and 5576 ± 1538 IE/g pancreas weight. An IU solution was made by adding iodixanol to University of Wisconsin solution and an IK solution was made by adding iodixanol to MK solution. The efficacy of the two solutions for islet isolation was compared. There were no significant differences between the two purification methods with regard to islet yield, survival rate, purity, score, or stimulation index. These results indicate that our isolation protocol produces efficient islet yields from prolonged cold-stored pancreas and that IU and IK solutions are equally useful for islet purification.

scholarly journals Comparison of Clostripain and Neutral Protease as Supplementary Enzymes for Human Islet Isolation

2018 ◽  
Vol 28 (2) ◽  
pp. 176-184 ◽  
Author(s):  
Heide Brandhorst ◽  
Paul R. Johnson ◽  
Johanna Mönch ◽  
Manfred Kurfürst ◽  
Olle Korsgren ◽  
...  
Keyword(s):  
Islet Transplantation ◽  
Stimulation Index ◽  
Effective Means ◽  
Insulin Response ◽  
Human Islets ◽  
Human Islet ◽  
Neutral Protease ◽  
Islet Isolation ◽  
Clinical Islet Transplantation ◽  
Glucose Challenge

Although human islet transplantation has been established as valid and safe treatment for patients with type 1 diabetes, the utilization rates of human pancreases for clinical islet transplantation are still limited and substantially determined by the quality and composition of collagenase blends. While function and integrity of collagenase has been extensively investigated, information is still lacking about the most suitable supplementary neutral proteases. The present study compared islet isolation outcome after pancreas digestion by means of collagenase used alone or supplemented with either neutral protease (NP), clostripain (CP), or both proteases. Decent amounts of islet equivalents (IEQ) were isolated using collagenase alone (3090 ± 550 IEQ/g), or in combination with NP (2340 ± 450 IEQ/g) or CP (2740 ± 280 IEQ/g). Nevertheless, the proportion of undigested tissue was higher after using collagenase alone (21.1 ± 1.1%, P < 0.05) compared with addition of NP (13.3 ± 2.2%) or CP plus NP (13.7 ± 2.6%). Likewise, the percentage of embedded islets was highest using collagenase only (13 ± 2%) and lowest adding NP plus CP (4 ± 1%, P < 0.01). The latter combination resulted in lowest post-culture overall survival (42.7 ± 3.9%), while highest survival was observed after supplementation with CP (74.5 ± 4.8%, P < 0.01). An insulin response toward glucose challenge was present in all experimental groups, but the stimulation index was significantly decreased using collagenase plus NP (2.0 ± 0.12) compared with supplementation with CP (3.16 ± 0.4, P < 0.001). This study demonstrates for the first time that it is possible to isolate significant numbers of human islets combining collagenase only with CP. The supplementation with CP is an effective means to substantially reduce NP activity, which significantly decreases survival and viability after culture. This will facilitate the manufacturing of enzyme blends with less harmful characteristics.

Evaluation of a Purified Enzyme Blend for the Recovery and Function of Canine Pancreatic Islets

1998 ◽  
Vol 7 (4) ◽  
pp. 365-372 ◽  
Author(s):  
Jonathan R.T. Lakey ◽  
Thomas J. Cavanagh ◽  
Michael A.J. Zieger ◽  
Mary Jo Wright
Keyword(s):  
Islet Transplantation ◽  
Stimulation Index ◽  
Insulin Response ◽  
Canine Model ◽  
Preclinical Model ◽  
Islet Isolation ◽  
Clinical Islet Transplantation ◽  
Type V ◽  
And Function

Recently developed technologies enabling the production of a reproducible, purified enzyme blend for optimal human pancreatic islet isolation has renewed interest in clinical islet transplantation. The canine model has been an ideal preclinical model for the development of islet transplantation protocols. As seen in other species, the application of crude collagenase for isolating canine islets resulted in highly variable islet yields, extensive islet fragmentation, and variable islet functionality. We compared the function of commercially available crude collagenases with that of Liberase™-CI purified enzyme blend for canine islet isolation. We also compared two manufacturing runs of Liberase-CI enzyme (lots 1 and 2) to demonstrate reproducibility of islet recovery and function. We report on the improved recovery and function of islets isolated using Liberase-CI enzyme. No difference in dog age, mean body weight, or pancreas weight were observed between the experimental groups. We observed a significantly higher postpurification recovery of islet equivalent number (IE) from pancreases processed using two lots of Liberase-CI enzyme (189 ± 20 × 103 IE, n = 4) and lot 2 (234 ± 39 × 103 IE, n = 7) than that obtained from pancreases processed with Sigma Type V (116.8 ± 27 × 103 IE, n = 5), Serva collagenase (49 ± 11.6 × 103 IE, n = 5, p < 0.05) or Boehringer–Mannheim (BM) Type P collagenase (85.4 ± 25 × 103 IE, n = 5, p < 0.05, ANOVA). No significant differences were observed in islet yield recovery from pancreases processed using the two production lots of Liberase-CI enzyme. Islet survival after 48 h in culture at 37°C was significantly higher from islets isolated using Liberase-CI enzyme (88 ± 3.7% survival) when compared to Sigma Type V (81.8 ± 3.3%), Serva (71.7 ± 2.8%), and BM Type P (77 ± 7.2%) (p < 0.05). Islet functional testing in vitro demonstrated islets isolated using crude collagenase had an increased insulin basal release and a reduced insulin stimulated response when compared with islets isolated using the two lots of Liberase-CI enzyme. The calculated stimulation index was 7.8 ± 1.7, 3.1 ± 0.6, and 4.8 ± 1.1 for Sigma Type V, Serva, and BM Type P isolated islets, respectively, compared to 15.7 ± 1.6 and 16.2 ± 1.9 for islets isolated with Liberase-CI enzyme production lots 1 and 2, respectively (p < 0.05). This evaluation demonstrates that a purified enzyme blend can significantly improve islet recovery and function. It also demonstrates the manufacturing reproducibility of Liberase-CI enzyme lots resulting in the isolation of canine islets with the same degree of efficacy. A blend of purified enzymes, specifically formulated for canine islet isolation, can consistently yield large numbers of islets that survive longer in culture and demonstrate an improved insulin response in vitro.

scholarly journals The Future of Clinical Islet Transplantation in the United States

2021 ◽  
Vol 5 (1) ◽  
Author(s):  
Michael F Knoll ◽  
Carmela A Knoll ◽  
Rita Bottino ◽  
Massimo Trucco ◽  
Suzanne Bertera ◽  
...  
Keyword(s):  
Quality Of Life ◽  
United States ◽  
Type 1 Diabetes ◽  
Islet Transplantation ◽  
Drug Administration ◽  
The United States ◽  
Clinical Islet Transplantation ◽  
The World

Clinical islet transplantation was first realized over four decades ago at the University of Minnesota. Autologous islet transplantation is now widely recognized as a treatment to prevent diabetes in patients after pancreas excision and is offered at major transplant centers throughout the United States and the world. Type 1 diabetes represents a much larger demographic in which islet transplantation may benefit patients. Allogeneic islet transplantation can now offer similar outcomes to pancreas transplantation in a subset of patients with labile type 1 diabetes with less risk than whole organ transplantation. It is recognized as a standard of care in nations around the world but not in the United States, despite the important developmental role US scientists and physicians have played. Early reports of islet transplantation focused on insulin independence that proved to diminish over time. However, regardless of insulin status, islet transplantation provides benefits ranging from improved quality of life to reduction in diabetic complications. A National Institutes of Health sponsored multi-center Phase 3 Clinical Trial (CIT-07) demonstrated safety and efficacy, although the Food and Drug Administration chose to consider islets as a biologic that requires licensure, which makes offering the procedure in the clinic very challenging. Until regulations can be brought into communion with international standards, allogeneic islet transplantation in the United States is unlikely to match international levels of success and once promising programs are left to wither on the vine. Food and Drug Administration approval would open the door for third party medical reimbursement and allow many patients the opportunity to enjoy better health and quality of life. Establishment of clinical islet transplantation for type 1 diabetes would lead to optimizations in procedures making it more efficacious and cost effective while offering support for ongoing islet xenotransplantation studies that could bring islet transplantation to even more patients.

Clinical islet transplantation in type 1 diabetes mellitus: results of Australia's first trial

2006 ◽  
Vol 184 (5) ◽  
pp. 221-225 ◽  
Author(s):  
Philip J O’Connell ◽  
Wayne J Hawthorne ◽  
Brian J Nankivell ◽  
Anita T Patel ◽  
Stacey N Walters ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Type 1 Diabetes ◽  
Type 1 Diabetes Mellitus ◽  
Islet Transplantation ◽  
Clinical Islet Transplantation

Excellence of the two-layer method (University of Wisconsin solution/perfluorochemical) in pancreas preservation before islet isolation

Surgery ◽  
1997 ◽  
Vol 122 (2) ◽  
pp. 435-442 ◽  
Author(s):  
Yasuki Tanioka ◽  
David E.R Sutherland ◽  
Yoshikazu Kuroda ◽  
Thomas R Gilmore ◽  
Tor C Asaheim ◽  
...  
Keyword(s):  
Islet Isolation ◽  
University Of Wisconsin Solution ◽  
University Of Wisconsin ◽  
Layer Method ◽  
Pancreas Preservation ◽  
Wisconsin Solution

NEW PANCREAS PRESERVATION STRATEGY FOR SUCCESSFUL CLINICAL ISLET TRANSPLANTATION

2008 ◽  
Vol 86 (Supplement) ◽  
pp. 142
Author(s):  
M Shimoda ◽  
H Noguchi ◽  
B Naziruddin ◽  
T Ikemoto ◽  
A Jackson ◽  
...  
Keyword(s):  
Islet Transplantation ◽  
Clinical Islet Transplantation ◽  
Pancreas Preservation

scholarly journals Pancreas Procurement and Preservation for Islet Transplantation: Personal Considerations

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Hirofumi Noguchi
Keyword(s):  
Islet Transplantation ◽  
Cooling System ◽  
Pancreatic Tissue ◽  
Diabetic Patients ◽  
Pancreatic Islet Transplantation ◽  
Ischemic Time ◽  
Trypsin Inhibition ◽  
Edmonton Protocol ◽  
Collagenase Inhibition ◽  
Type 1 Diabetic Patients

Pancreatic islet transplantation is a promising option for the treatment of type 1 diabetic patients. After the successful demonstration of the Edmonton protocol, islet transplantation has advanced significantly on several fronts, including improved pancreas procurement and preservation systems. Since we frequently use pancreata from donors after cardiac death in Japan,we have applied thein situregional organ cooling system for pancreas procurement to reduce the warm ischemic time. To reduce the apoptosis of pancreatic tissue during cold preservation, we have applied the ductal injection of preservation solution. For pancreas preservation, we use modified Kyoto solution, which is advantageous at trypsin inhibition and less collagenase inhibition. In this paper, we show pancreas procurement and preservation in our group for islet transplantation.

scholarly journals Animal Models of Diabetes Mellitus for Islet Transplantation

2012 ◽  
Vol 2012 ◽  
pp. 1-11 ◽  
Author(s):  
Naoaki Sakata ◽  
Gumpei Yoshimatsu ◽  
Haruyuki Tsuchiya ◽  
Shinichi Egawa ◽  
Michiaki Unno
Keyword(s):  
Diabetes Mellitus ◽  
Animal Models ◽  
Islet Transplantation ◽  
Diabetic Animal ◽  
Islet Isolation ◽  
Loss Of Function ◽  
Severe Diabetes ◽  
Animal Models Of Diabetes ◽  
Type 1 Dm

Due to current improvements in techniques for islet isolation and transplantation and protocols for immunosuppressants, islet transplantation has become an effective treatment for severe diabetes patients. Many diabetic animal models have contributed to such improvements. In this paper, we focus on 3 types of models with different mechanisms for inducing diabetes mellitus (DM): models induced by drugs including streptozotocin (STZ), pancreatomized models, and spontaneous models due to autoimmunity. STZ-induced diabetes is one of the most commonly used experimental diabetic models and is employed using many specimens including rodents, pigs or monkeys. The management of STZ models is well established for islet studies. Pancreatomized models reveal different aspects compared to STZ-induced models in terms of loss of function in the increase and decrease of blood glucose and therefore are useful for evaluating the condition in total pancreatomized patients. Spontaneous models are useful for preclinical studies including the assessment of immunosuppressants because such models involve the same mechanisms as type 1 DM in the clinical setting. In conclusion, islet researchers should select suitable diabetic animal models according to the aim of the study.

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