Notice of forthcoming erratum

Porcine Islet ◽

University Of Wisconsin ◽

Published Paper ◽

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

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

The Scientific World JOURNAL ◽

10.1100/tsw.2003.107 ◽

2003 ◽

Vol 3 ◽

pp. 1154-1159 ◽

Cited By ~ 14

Author(s):

Michael P.M. Van der Burg ◽

John M. Graham

Keyword(s):

Adverse Effects ◽

Human Islet ◽

Porcine Pancreas ◽

Isolated Pancreatic Islets ◽

Porcine Islet ◽

University Of Wisconsin ◽

Ficoll Gradient ◽

The University ◽

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.

No Porcine Islet Loss During Density Gradient Purification in a Novel Iodixanol in University of Wisconsin Solution

Transplantation Proceedings ◽

10.1016/s0041-1345(97)01308-0 ◽

1998 ◽

Vol 30 (2) ◽

pp. 362-363 ◽

Cited By ~ 24

Author(s):

M.P.M van der Burg ◽

I Basir ◽

E Bouwman

Keyword(s):

Density Gradient ◽

Porcine Islet ◽

University Of Wisconsin ◽

Density Gradient Purification ◽

Porcine Islet Preservation During Isolation in University of Wisconsin Solution

Transplantation Proceedings ◽

10.1016/s0041-1345(97)01307-9 ◽

1998 ◽

Vol 30 (2) ◽

pp. 360-361 ◽

Cited By ~ 6

Author(s):

M.P.M. van der Burg ◽

I. Basir ◽

R.P. Zwaan ◽

E. Bouwman

Keyword(s):

Porcine Islet ◽

University Of Wisconsin ◽

Cell Preservation in University of Wisconsin Solution during Isolation of Canine Islets of Langerhans

Cell Transplantation ◽

10.1177/096368979400300408 ◽

1994 ◽

Vol 3 (4) ◽

pp. 315-324 ◽

Cited By ~ 7

Author(s):

Michael P.M. Van Der Burg ◽

Onno R. Guicherit ◽

Marijke Frölich ◽

Frans A. Prins ◽

Jan Anthonie Bruijn ◽

...

Keyword(s):

Density Gradient ◽

Acinar Cells ◽

Cell Swelling ◽

Type I ◽

Islet Isolation ◽

Isolation And Purification ◽

Density Separation ◽

University Of Wisconsin ◽

Allogeneic islet transplantation in Type I diabetic patients is considerably hampered by the variable outcome of islet isolation and purification. After collagenase digestion of the pancreas, islet isolation is traditionally performed under hypothermic conditions in physiological solutions such as Hanks and RPMI. The University of Wisconsin solution (UWS) has been shown superior for hypothermic preservation of the pancreas. We, therefore, compared the UWS and RPMI for canine islet isolation and subsequent purification in either a conventional hyperosmotic density gradient of dextran in Hanks, or a novel normosmotic density gradient of Percoll in UWS. The isolation solution did not affect islet yield before purification (51% of the native islet mass). Loss of amylase (30%) and swelling of the acinar cells were observed in RPMI. In contrast, no loss of amylase and slight shrinkage of the acinar cells were observed in the UWS. Cell swelling affected the density separation and viability of the cells. Dextran density separation resulted in a 15% purity and 41% recovery of the islets isolated in RPMI, as compared to a 93% purity and 52% recovery of islets isolated in UWS. Percoll density separation improved the purity (99%) and recovery (74%) of islets isolated in UWS. Islets isolated in UWS demonstrated a superior basal and glucose stimulated insulin release during perifusion. Electron microscopy demonstrated a well-preserved islet ultrastructure after isolation in both solutions — except for slightly swollen mitochondria after isolation in RPMI. Autotransplantation of islets in pancreatectomised dogs was successful both after isolation in UWS and RPMI. We conclude that prevention of cell swelling during isolation and purification in the UWS resulted in an improved yield of viable and consistent virtually pure islets. Prevention of cell swelling during islet isolation should facilitate the analysis and control of other factors affecting outcome in man.

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

Cell Medicine ◽

10.1177/2155179018775071 ◽

2018 ◽

Vol 10 ◽

pp. 215517901877507 ◽

Cited By ~ 1

Author(s):

Yoshiki Nakashima ◽

Chika Miyagi-Shiohira ◽

Nana Ebi ◽

Eri Hamada ◽

Yoshihito Tamaki ◽

...

Keyword(s):

Survival Rate ◽

Pancreatic Islets ◽

Stimulation Index ◽

Islet Isolation ◽

Porcine Islet ◽

University Of Wisconsin ◽

Purification Methods ◽

Isolation Performance ◽

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.

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

Journal of Clinical Medicine ◽

10.3390/jcm8101561 ◽

2019 ◽

Vol 8 (10) ◽

pp. 1561 ◽

Cited By ~ 3

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 ◽

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.