Iodixanol-UWS Purification and Culture of Pig Islets Xeno Transplants • Cell Transplant Society 1999 • @1MEMO 20250202


Iodixanol-UWS Purification and Culture of Pig Islets Xeno Transplants • Cell Transplant Society 1999 • Michel van der Burg • Miracles.Media • @1MEMO 20250202

Slideshow (digitally remastered) oral presentation Michel van der Burg, Cell Transplant Society congress 1999 in Montreux, Switzerland (1)

The Leiden Islet Laboratory developed a novel Iodixanol-UWS density gradient for purification of the islets of Langerhans from the pancreas in the difficult pig model.

Pig islet isolation from the pancreas of slaughterhouse sows was markedly improved by keeping the islets in the UW donor organ preservation solution (UWS) both during the isolation of islets (using the Liberase enzyme blend) and purification of the islets using our novel Iodixanol-UWS density gradient.

Xeno transplantation of these 1-day-cultured pig islets restored normal blood sugar levels in diabetic mice.

PDF slideshow oral presentation Michel van der Burg, Cell Transplant Society congress 1999 in Montreux, Switzerland

Iodixanol-UWS Purification and Culture of Pig Islets Xeno Transplants • Cell Transplant Society 1999 • Michel van der Burg • Miracles.Media • @1MEMO 20250202_2 • TakeNode 8bd79256-78b2-4d8e-8d48-6da41102dc2d • Download file available below 1MEMO_20250202_2

Notes

1) Van der Burg MPM, Rijkelijkhuizen JKRA, Zwaan RP, Bouwman E. Adult pig islet recovery during Liberase isolation, OptiPrep purification and culture for transplantation in nude mice. 4th International congress of the Cell Transplant Society, Montreux (Switzerland) March 21–24, 1999 .

2) Van der Burg MPM, Rijkelijkhuizen JKRA, Zwaan RP, Bouwman E. Adult pig islet recovery during Liberase isolation, OptiPrep purification and culture for transplantation in nude mice (Abstract). Cell Transplant 1999; 8: 185. Download abstract, available below 1MEMO_20250202_3

Citation info : Iodixanol-UWS Purification and Culture of Pig Islets Xeno Transplants • Cell Transplant Society 1999 • Michel van der Burg • Miracles.Media • @1MEMO 20250202 • TakeNode f9d1fdc5-21e4-4932-b995-4c97639945fe • URL michelvanderburg.com/2025/02/02/

Fresh vs Cultured Pig Islets for Xenografts • NTV Bootcongres 1998 • @1MEMO 20250201

Fresh vs Cultured Pig Islets for Xenografts • NTV Bootcongres 1998 • Michel van der Burg • Miracles.Media • @1MEMO 20250201_1 • TakeNode 71d39e7e-ab5e-42b6-8cf2-a17d464ce132


Van der Burg MPM, Rijkelijkhuizen JKRA, Zwaan RP, Bouwman E. Culture of isolated pig islets pre-xenotransplantation greatly improves the quality and survival of the graft (Kweek van geïsoleerde varkenseilandjes pre-xenotransplantatie verbetert aanzienlijk de kwaliteit en overleving van het transplantaat). Tenth Congress of the Dutch Transplantation Society (Bootcongres 1998), Kerkrade (The Netherlands) April 21–23, 1998.

Download (pdf) NTV bulletin (abstract) & poster , file 1MEMO_20250201

EN – Abstract

PRE-TRANSPLANT SHORT-TERM CULTURE OF ISOLATED PIG ISLETS MARKEDLY IMPROVES THE QUALITY AND SURVIVAL OF THE XENOGRAFT
Van der Burg MPM,* Rijkelijkhuizen JKRA, Zwaan RP, and Bouwman E, Leiden University Medical Centre, Leiden, The Netherlands.

Pre-transplant culture of isolated islets reduces the graft’s immunogenicity in various models. Little is known, however, on the effect of culture in pig islet xenotransplantation — probably, because porcine islets have been found difficult to culture. Because pilot transplants of freshly-isolated pig islets in strongly immunosuppressed rats resulted in primary non function, we first studied 1-7 day culture of the isolated pig islets at 37°C in RPMI plus 10% porcine serum. Islets were isolated from large sows (n =6) by an improved method, resulting in intact islets (size 185±16 µm) and no loss during purification, yielding 2448 islets (IEQs)/g with a >95% purity and 90±2% viability as assessed by acridine orange – propidium iodide (AOPI) staining. During culture, however, islet recovery was 24±9% at day 1 and 17±6% at day 7 (NS vs day 1). In order to delineate whether the culture conditions or the quality of freshly isolated islets caused the islet loss during culture, we compared graft survival in nude mice at 1 mo after transplantation under the kidney capsule of ~2500 fresh islets in normoglycemic recipients or ~1000 cultured islets in STZ-diabetic (>20 mM) recipients. After transplanting fresh islets, histology of the kidneys — sectioned every 500 µm — demonstrated substantial scarring and (near-)absence of islets. Cultured islet transplants, by contrast, rendered 5/6 recipients normoglycemic, and showed a substantial mass of well-preserved islets with little scarring at the grafts’ site. Thus, viability by AOPI-staining of fresh islets poorly predicts the survival in vivo and in vitro. A non-immune mediated disintegration of part of the fresh islets may substantially reduce the functional capacity of the graft, both direct by lowering the effective islet dose, and indirect first because scarring may hamper the engraftment of viable tissue, and second because the cellular debris most probably will attract macrophages and induce the release of harmful cytokines.

Citation info : Fresh vs Cultured Pig Islets for Xenografts • NTV Bootcongres 1998 • Michel van der Burg • Miracles.Media • @1MEMO 20250201 • URL michelvanderburg.com/2025/02/01/

Iodixanol-UWS Purification for Porcine Islet Transplantation • AIDSPIT 1998 • @1MEMO 20250131

Iodixanol-UWS Purification for Porcine Islet Transplantation • AIDSPIT 1998 • Michel van der Burg • Miracles.Media • @1MEMO 20250131

Slideshow (digitally remastered) of first talk on pancreatic islet xeno-transplantation following adult pig islet purification in our novel Iodixanol-UWS density gradient. Presented at the 17th Workshop of the AIDSPIT Study Group in Igls, Austria, Tuesday, Jan 27, 1998 (1, 2).

Isolation of porcine pancreatic islets (from the pancreas of large sows) was markedly improved by keeping the islets in the UW donor organ preservation solution (UWS) both during the isolation of islets (using the Liberase enzyme blend) and purification of the islets using a novel Iodixanol-UWS density gradient.
Transplantation of the pig islets (after 1 day storage in culture solution) restored normal blood sugar levels in diabetic mice.


20250131 1

Notes

1) Van der Burg MPM, Zwaan RP, Bouwman E. Markedly improved outcome of adult porcine islet isolation, purification, and culture using Liberase-PI versus collagenase-P, and a novel gradient of Optiprep in University of Wisconsin solution. 17th Workshop of the Study Group on Artificial Insulin Delivery Systems, Pancreas and Islet Transplantation (AIDSPIT) of the European Association for the Study of Diabetes (EASD), Igls (Austria) January 25—27,1998

2) Van der Burg MPM, Zwaan RP, Bouwman E. Markedly improved outcome of adult porcine islet isolation, purification, and culture using Liberase-PI versus collagenase-P, and a novel gradient of Optiprep in University of Wisconsin solution (Abstract). Horm Metab Res 1998; 30(1): A23 . DOI: 10.1055/s-2007-978834 .
Download abstract 1MEMO_20250131_2 below :

Citation info : Iodixanol-UWS Purification for Porcine Islet Transplantation • AIDSPIT 1998 • Michel van der Burg • Miracles.Media • @1MEMO 20250131 • TakeNode 1f185515-7aeb-44ac-b721-2cec76df67b7 • URL michelvanderburg.com/2025/01/31/

Innovative Islet Purification in Iodixanol-UWS • IPITA 97 • @1MEMO 20250128

No porcine islet loss during density gradient purification in a novel iodixanol in University of Wisconsin solution. First talk on our introduction of the innovative Iodixanol-UWS density gradients for islet purification , showing no islet loss in the difficult model of pig islet isolation — presented at the 6th IPITA congress in Milan (Italy) , Thursday, 25th september 1997 (1).

Background

In 1995 the Leiden Islet Lab had moved into the Surgery Lab in the new LUMC building, and the islet team had grown, focussing on xenotransplantation. With more people isolating pig islets, more convenient islet isolation solutions were tried.

Both Percoll-UWS (2) and Histopaque-UWS (3) were being used from 1992 in first series of islet isolations from juvenile pig pancreases.

August 1995, I started designing the clinical promising islet purification solution of Iodixanal in UW-Solution (UWS). I mixed Optiprep – a 60 % solution of iodixanol in water – with a 2-times concentrated UWS, to prepare this density gradient solution. The first standard operation procedure of this Iodixanol-UWS gradient for our pig islet isolation research was completed November 1995.

This purification solution was a logical follow-up of our highly succesful Percoll-UWS density gradient (2) — a similar solution of impermeants (that do not readily enter cells) with similar osmolarity (close to that of the tissue) , more practical though, and with the important advantage that iodixanol is clinical tested.

July 1996, I introduced the Iodixanol-UWS gradient to replace the Percoll-UWS gradient for large scale pancreatic islet research in this difficult model of islet isolation from juvenile pig pancreases.

The work was presented September 1997. First a poster was presented that month at the Xenotransplantation Congress in Nantes , France (4). Next, two talks followed at the 6th Congress of the International Pancreas and Islet Transplant Association (IPITA) in Milan (Italy).

Here the (remastered) slideshow of that first talk at the 6th IPITA Congress in Milan, presented Thursday, 25th september 1997 (1) . The slideshow of the 2nd talk , presented at the last day of that congress, was posted yesterday (5).

Notes

1) Van der Burg MPM, Basir I, Bouwman E. No porcine islet loss during density gradient purification in a novel iodixanol in University of Wisconsin solution. Sixth Congress of the Int. Pancreas and Islet Transplant Association (IPITA), Milan (Italy) Sept. 24–27, 1997.

2) Percoll UWS Purification Pancreatic Islets • ESOT 1991 Talk • Michel van der Burg • Miracles.Media • @1MEMO 20250114 • URL michelvanderburg.com/2025/01/14/

3) Basir I, van der Burg MPM, Scheringa M, Töns A, Bouwman E. Improved outcome of pig islet isolation by Pefabloc inhibition of trypsin. Transplant Proc. 1997 Jun;29(4):1939-41. doi: 10.1016/s0041-1345(97)00168-1. PMID: 9193462

4) Van der Burg MPM, Basir I, Bouwman E. Non-traumatic porcine islet density gradient purification in a novel iodixanol in University of Wisconsin solution. The 4th Int. Congress for Xenotransplantation, Nantes (France) Sept. 7–11, 1997. Online in : Purification Islets in Iodixanol-UWS • Leiden Start 1996 • Michel van der Burg • Miracles.Media • @1MEMO 20250126 • URL michelvanderburg.com/2025/01/26/

5) Van der Burg MPM, Basir I, Zwaan RP, Bouwman E. Porcine islet preservation during isolation in University of Wisconsin solution. Sixth Congress of the Int. Pancreas and Islet Transplant Association (IPITA), Milan (Italy) Sept. 24–27, 1997. Online in : Porcine Islet Preservation In UWS • IPITA 97 • Michel van der Burg • Miracles.Media • @1MEMO 20250127• URL michelvanderburg.com/2025/01/27/

Milan Papers • @1MEMO 20250205 • Manuscripts (Preprints) and Abstracts copies of the presentations at the IPITA 1997 meeting in Milan (Italy). PDF downloads at michelvanderburg•com/2025/02/05/

Citation info : Innovative Islet Purification in Iodixanol-UWS • IPITA 97 • Michel van der Burg • Miracles.Media • @1MEMO 20250128 • TakeNode • URL michelvanderburg.com/2025/01/28/

Porcine Islet Preservation In UWS • IPITA 97 • @1MEMO 20250127

August 1995, I started designing the clinical promising islet purification solution of Iodixanal in UW-Solution (UWS). I mixed Optiprep – a 60 % solution of iodixanol in water – with a 2-times concentrated UWS, to prepare this density gradient solution. The first standard operation procedure of this Iodixanol-UWS gradient for our pig islet isolation research was completed November 1995.

This purification solution was a logical follow-up of our highly succesful Percoll-UWS density gradient — a similar solution of impermeants (that do not readily enter cells) with similar osmolarity (close to that of the tissue) , more practical though, and with the important advantage that iodixanol is clinical tested.

The Leiden Islet Lab had just moved into the Surgery Lab in the new LUMC building, and the islet team had grown, focussing on xenotransplantation. With more people isolating pig islets, more convenient solutions were tried.

Both Percoll-UWS and Histopaque-UWS were being used from 1992 for small scale islet isolations from juvenile pig pancreases.

July 1996, I introduced the Optiprep-UWS gradient to replace our Percoll-UWS gradient for larger scale porcine islet isolations.

The work was first presented September 1997, both in a poster presented at the Xenotransplantation Congress in Nantes , France (1), and later that month in two talks at the 6th Congress of the International Pancreas and Islet Transplant Association (IPITA) in Milan (Italy). The slideshow of the first talk (2) will be online at this site soon.
Here the remastered slideshow of the 2nd talk at the 6th IPITA Congress in Milan, presented Saturday, 27th September, 1997 (3).

Notes

1) Van der Burg MPM, Basir I, Bouwman E. Non-traumatic porcine islet density gradient purification in a novel iodixanol in University of Wisconsin solution. The 4th Int. Congress for Xenotransplantation, Nantes (France) Sept. 7–11, 1997. Online in : Purification Islets in Iodixanol-UWS • Leiden Start 1996 • Michel van der Burg • Miracles.Media • @1MEMO 20250126 • URL michelvanderburg.com/2025/01/26/

2) Van der Burg MPM, Basir I, Bouwman E. No porcine islet loss during density gradient purification in a novel iodixanol in University of Wisconsin solution. Sixth Congress of the Int. Pancreas and Islet Transplant Association (IPITA), Milan (Italy) Sept. 24–27, 1997.

3) Van der Burg MPM, Basir I, Zwaan RP, Bouwman E. Porcine islet preservation during isolation in University of Wisconsin solution. Sixth Congress of the Int. Pancreas and Islet Transplant Association (IPITA), Milan (Italy) Sept. 24–27, 1997.

Milan Papers • @1MEMO 20250205 • Manuscripts (Preprints) and Abstracts copies of the presentations at the IPITA 1997 meeting in Milan (Italy). PDF downloads at michelvanderburg•com/2025/02/05/

Update 20250128 – update intro design timeline

Update 20250205 – added Milan papers

Citation info : Porcine Islet Preservation In UWS • IPITA 97 • Michel van der Burg • Miracles.Media • @1MEMO 20250127 • TakeNode 6d6fca2e-014c-40cf-8110-725b267c0b0e • URL michelvanderburg.com/2025/01/27/

Leiden Islet Isolation Chamber • @1MEMO 20250116

Leiden Islet Isolation Chamber • Michel van der Burg • Miracles.Media • @1MEMO 20250116 • TakeNode 398303fe-928a-4c53-839e-9dde0e7d8ec6

Following a working visit February ’92 studying pig islet isolation at the ‘Fondation Transplantation’ in Strasbourg -Hautepierre, France (1) , I designed this islet isolation chamber March 1992 for use in our Leiden Islet Laboratory , together with Gerard Verschagen , head of the precision mechanics workshop of the Laboratory of Physiology of the University Hospital Leiden (AZL , now LUMC), who manufactured this and other of our islet isolation devices.

The chamber was designed with support for use together with the Retsch Vibro Sieving Machine , purchased earlier for our Leiden Islet Laboratory, a vibrating sieving machine I had used in the 70s for isolation cell nuclei (2). The design was based on the Ricordi chamber published 4 years earlier (3).

The drawing was originally designed and produced on a Macintosh computer March 1992, using the MacDraw II application. That Mac file was remastered today in LibreOffice Vanilla, by straightening most of the lines that are slanted after opening in LibreOffice and correcting the font changes. So , not accurate now probably … just for illustration the history of progress in our Leiden Islet Laboratory, and love for art ;)

Notes

1) Xeno Islets • Michel van der Burg • Miracles.Media • @1MEMO 20250113 • URL michelvanderburg.com/2025/01/13/

2) Mennes AM, Bouman H, Van der Burg MPM, Libbenga KR. RNA synthesis in isolated
tobacco callus nuclei, and the influence of phytohormones. Plant Sci Lett 1978; 13: 329– 339, ISSN 0304-4211, https://doi.org/10.1016/0304-4211(78)90210-9.

3) Piemonti L, Pileggi A. 25 Years Of The Ricordi Automated Method For Islet Isolation. CellR4 Repair Replace Regen Reprogram. 2013;1(1):e128. PMID: 30505878; PMCID: PMC6267808. URL https://www.cellr4.org/article/128

Citation info : Leiden Islet Isolation Chamber • Michel van der Burg • Miracles.Media • @1MEMO 20250116 • URL michelvanderburg.com/2025/01/16/

Xeno Islets • @1MEMO 20250113

Pig Islet Isolation, Fondation Transplantation, Strasbourg 1992 • Michel van der Burg • Miracles.Media • @1MEMO 20250113_1 • TakeNode b1f66781-cf63-4e64-a7b3-24cbdf5c107f

For the xenotransplantation research we started in 1991 at our Leiden Surgery Department, I studied pig islet isolation during a working visit February 1992, at the ‘Fondation Transplantation’ in Strasbourg-Hautepierre, France — here watching our student Vincent Nieuwenhuijs (left) at work in the international team – with Agnaldo Soares I guess – using the Ricordi method for islet isolation — and presented the recent Leiden islet work (1).

Notes

1) Van der Burg MPM, Gooszen HG. Islet preservation using UW, and islet function after
autotransplantation in dogs. Fondation Transplantation, CHU HautePierre Strasbourg
(France) February 18, 1992.

Update 20250124 – Paper reference

Citation info : Xeno Islets • Michel van der Burg • Miracles.Media • @1MEMO 20250113 • URL michelvanderburg.com/2025/01/13/