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 A Comparison of Pancreatic Islet Purification using Iodixanol with University of Wisconsin Solution and with Na-Lactobionate and Histidine Solution

Cell Medicine ◽  
2018 ◽  
Vol 10 ◽  
pp. 215517901877507 ◽  
Author(s):  
Yoshiki Nakashima ◽  
Chika Miyagi-Shiohira ◽  
Nana Ebi ◽  
Eri Hamada ◽  
Yoshihito Tamaki ◽  
...  
Keyword(s):  
Survival Rate ◽  
Pancreatic Islets ◽  
Stimulation Index ◽  
Islet Isolation ◽  
University Of Wisconsin Solution ◽  
Porcine Islet ◽  
University Of Wisconsin ◽  
Purification Methods ◽  
Isolation Performance ◽  
Wisconsin Solution

Purification of pancreatic islets is an important step in islet isolation for islet transplantation. In this study, to investigate how a solution composed mainly of Na-lactobionate and histidine (HL) influences the purification of islets, iodixanol was added to a purified solution for porcine islet isolation. A solution (IU) made by adding iodixanol to University of Wisconsin solution and a solution (IHL) made by adding iodixanol to HL solution were used to evaluate the islet isolation performance. We noted no significant differences between the two purification methods with regard to the islet yield, survival rate or purity, score, or stimulation index. These results show that IHL solution is as useful as IU solution for islet purification.

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.

Pancreas Preservation in Modified Histidine-lactobionate Solution Is Superior to That in University of Wisconsin Solution for Porcine Islet Isolation

2021 ◽  
Vol Publish Ahead of Print ◽  
Author(s):  
Mayuko Sakai-Yonaha ◽  
Chika Miyagi-Shiohira ◽  
Kazuho Kuwae ◽  
Yoshihito Tamaki ◽  
Kai Nishime ◽  
...  
Keyword(s):  
Islet Isolation ◽  
University Of Wisconsin Solution ◽  
Porcine Islet ◽  
University Of Wisconsin ◽  
Pancreas Preservation ◽  
Wisconsin Solution

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

scholarly journals Iodixanol Density Gradient Preparation in University of Wisconsin Solution for Porcine Islet Purification

2003 ◽  
Vol 3 ◽  
pp. 1154-1159 ◽  
Author(s):  
Michael P.M. Van der Burg ◽  
John M. Graham
Keyword(s):  
Adverse Effects ◽  
Human Islet ◽  
Porcine Pancreas ◽  
Isolated Pancreatic Islets ◽  
University Of Wisconsin Solution ◽  
Porcine Islet ◽  
University Of Wisconsin ◽  
Ficoll Gradient ◽  
The University ◽  
Wisconsin Solution

Previously published as Graham, J.M. (2002) Purification of Islets of Langerhans from porcine pancreas. TheScientificWorldJOURNAL 2, 1657–1661. ISSN 1537-744X; DOI 10.1100/tsw.2002.847.Generally, prior to the purification of isolated pancreatic islets, the collagenase-digested tissue is incubated in the University of Wisconsin solution (UWS; ~320 mOsm) for osmotic stabilization to preserve or improve the density differences between islets and acinar fragments. The adverse effects arising from the subsequent pelleting and resuspension of the islets in a second, different (often highly hyperosmotic) purification solution are avoided in the protocol described here; preparation of the purification medium is simply achieved by mixing the UWS preincubated islets with a second UWS containing the inert impermeant iodixanol. Flotation of the islets isolated from juvenile porcine pancreases through this mildly hypertonic (~380 mOsm) gradient of iodixanol-UWS achieves a much higher recovery of islets of an improved viability than the customary method using a Ficoll gradient. The method has been extended to human islet purification.

The Effect of UW Solution and Its Components on the Collagenase Digestion of Human and Porcine Pancreas

1995 ◽  
Vol 4 (6) ◽  
pp. 615-619 ◽  
Author(s):  
Harold H. Contractor ◽  
Paul R. V. Johnson ◽  
David R. Chadwick ◽  
Gavin S. M. Robertson ◽  
Nicholas J. M. London
Keyword(s):  
Cold Storage ◽  
Species Difference ◽  
Human Pancreas ◽  
Islet Isolation ◽  
Porcine Pancreas ◽  
Uw Solution ◽  
Porcine Islet ◽  
University Of Wisconsin ◽  
Collagenase Digestion ◽  
Tissue Dissociation

University of Wisconsin (UW) solution is used extensively as a cold storage solution during the procurement and transport of the pancreas prior to islet isolation. However, it has been observed that UW inhibits the collagenase digestion phase of human but not porcine islet isolation, resulting in poor islet yields and islets of poor viability. The aim of this study was, therefore, to confirm this species difference and to determine which components of UW are responsible for the inhibition in the human. In the initial experiment, blocks of human and porcine pancreas (n = 7) were incubated in test tubes containing collagenase at a concentration of 4 mg/mL at 37°C dissolved in 4 mL of either Hanks' solution or UW. Every 5 min the tubes were manually shaken and the degree of tissue dissociation scored on a scale of + and +++. Our results confirm the inhibition of collagenase digestion in the human but not the pig. Using the same methodology, we then investigated the components of UW that were causing the observed inhibition in the human pancreas (n = 7). This time the collagenase was dissolved in individual or combinations of UW components. Using Hank's as a control, the results were then expressed as a median ratio. The components found to be most inhibitory were magnesium, the Na+/K+ ratio, hydroxyethyl starch (HES), and adenosine. Allopurinol in combination with either lactobionate or glutathione was markedly inhibitory (i.e., median ratio 1.8 and 1.9, respectively). The most inhibitory solution tested was a combination of the three components raffinose, glutathione, and lactobionate (median ratio 2.1). This combination was almost as inhibitory as UW itself (median ratio 2.7). These findings are essential for the development of effective cold-storage solutions for the human pancreas that do not inhibit the subsequent collagenase digestion phase of islet isolation.

scholarly journals Notice of forthcoming erratum

2003 ◽  
Vol 3 ◽  
pp. 826-826
Keyword(s):  
Density Gradient ◽  
Islets Of Langerhans ◽  
Porcine Pancreas ◽  
University Of Wisconsin Solution ◽  
Porcine Islet ◽  
University Of Wisconsin ◽  
Published Paper ◽  
Wisconsin Solution

The recently published peer-reviewed protocol article entitled Purification of Islets of Langerhans from Porcine Pancreas by John M. Graham (TheScientificWorldJOURNAL (2002) 2, 1657�1661, ISSN 1537-744X; DOI 10.1100/tsw.2002.847) is being revised. TheScientificWorldJOURNAL will shortly publish the article as an erratum entitled Iodixanol Density Gradient Preparation in University of Wisconsin Solution for Porcine Islet Purification. Links between the published paper and erratum will be made to facilitate cross-referencing.22 August 2003TheScientificWorldJOURNAL

scholarly journals Evaluation of Islet Purification Methods for Making a Continuous Density Gradient and Loading Tissue

Cell Medicine ◽  
2018 ◽  
Vol 10 ◽  
pp. 215517901773309 ◽  
Author(s):  
Nana Ebi ◽  
Chika Miyagi-Shiohira ◽  
Eri Hamada ◽  
Yoshihito Tamaki ◽  
Mariko Masamoto ◽  
...  
Keyword(s):  
Density Gradient ◽  
Stimulation Index ◽  
High Yield ◽  
Islet Isolation ◽  
Pancreatic Islet Transplantation ◽  
Porcine Pancreas ◽  
Purification Method ◽  
Continuous Density ◽  
Yield Rate ◽  
Purification Methods

Islet purification is one of the most important steps of islet isolation for pancreatic islet transplantation. We previously reported that a purification method using large plastic bottles effectively achieved a high yield of islets from porcine pancreas. In this study, we evaluated the methods for making a continuous density gradient and loading tissue. One method involved loading digested tissue on top of a continuous density gradient (top loading). The other method involved mixing digested tissue with low-density solution and then making a continuous gradient (mixed loading). There were no significant differences between the 2 purification methods in terms of the islet yield, rate of viability or purity, score, or in the stimulation index after purification. Furthermore, there were no marked differences in the attainability or suitability of posttransplantation normoglycemia. Our study shows the equivalency of these 2 methods of islet purification.

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