Oral Presentation Slide 19 ~ Pure Islets Isolated in UW Solution • Miracles.Media • 20241229_1 • TakeNode ce420e5a-1df7-4e4f-bcb6-6831434ffac7
In November 1989, I had my first chance for a presentation at a major international conference, the 4th ESOT meeting in Barcelona, of a paper centered on the rationale for employing the University of Wisconsin (UW) organ preservation solution in pancreatic islet isolation.
Venue 4th Congress of the European Society for Organ Transplantation, Barcelona (Spain) November 1–4, 1989 • Michel van der Burg • Miracles.Media • 20241224 • TakeNode 103c4c3c-5d36-4111-a5f8-f60e81c5db4d
This innovative islet isolation work with the UW-Solution had been accepted as a poster presentation for the 4th ESOT meeting (1). I also took the opportunity to present this new work during a talk on previous isolation work (2,3) at this conference.
Barcelona Nov 1989 4th ESOT • Talk by Michel van der Burg • Miracles.Media • 20241229_2 • Photo : Grup Jaume Muntaner (Barcelona) • TakeNode 96957e2f-1462-48e1-aca1-4745198d9a54
‘Behind the scenes’ I had first remade my lost poster at the congress venue (1), and then I rewrote my speech in the hotel room on the older research (2) updated with the more recent work with the UW-Solution, and including the outcome of our first islet transplantations.
Barcelona 89 • Handout 4th ESOT Talk
Below a scan of that handout of the final speech, written out at full lenght in a notepad from a recent working visit in Minneapolis (4).
Barcelona 89 • Handout 4th ESOT Talk • Michel van der Burg • Miracles.Media • 20241229_3 • TakeNode f8595a82-fba5-4a89-a01f-4c37976d06d7 • PDF FILE 20241229_3 copy is available here
( finishing speech first part on our previous islet isolation methods using the conventional RPMI solution – a solution originally designed for tissue culture, not for cold storage of organs )
Slide 18
Image Slide 18 (scan of print) • Best (Not Usual) Purity Islets RPMI Series • Miracles.Media • 20241229_4 • TakeNode a6afbf29-b3e6-41d0-abd0-07471303be15
This slide shows one of the purest preparations obtained, with an estimated purity of 65% islets. • On the average however a purity of only 30% was obtained.
Recently we adopted an entirely new approach to islet isolation. • Since UW, the new organ cold storage solution has been shown to allow long-term cold storage of the canine pancreas; • and, since islet isolation too is largely performed in the cold; • we tested this preservation solution as the isolation medium, throughout the isolation procedure in another 6 dogs; • as compared to the commonly used solutions like RPMI tissue culture medium.
Slide 19
Image Slide 19 (scan of print) – Usual Purity Islets UWS Series • Miracles.Media • 20241229_5 • TakeNode dacd3b99-5958-44fe-85ed-6abd55db2676
This slide shows the final dithizone stained preparation after dextran purification of UW-isolated islets. • UW did not affect islet yield • however, UW markedly improved purity : • acinar tissue is virtually absent.
2) Van der Burg MPM, Gooszen HG, Field MJ, Scherft JP, Terpstra JL, Van de Woude FT, Guicherit OR, Frölich M, Bruijn JA. Comparison of current islet isolation techniques in dogs. 4th Congress of the European Society for Organ Transplantation, Barcelona (Spain) November 1–4, 1989. Oral presentation. (1)
3) Van der Burg MPM, Gooszen HG, Field MJ, Scherft JP, Terpstra JL, van de Woude FJ, Guicherit OR, Frölich M, Bruijn JA. Comparison of current islet isolation techniques in dogs. Transplant Proc. 1990 Aug;22(4):2044-5. PMID: 1697121. This paper appears to be not readily available online – a reprint copy is available at this site (1)
4) Working visit Sep – Oct 1989, joining the pancreatic islet isolation team of the pioneer transplant surgeon David Sutherland at the University of Minnesota in Minneapolis.
Citation info : Pancreatic Islet Isolation in UW Solution & Transplantation • ESOT 1989 Talk • Michel van der Burg • Miracles.Media • @1MEMO 20241229 • URL michelvanderburg.com/2024/12/29/
As reported in our poster and the manuscript delivered in Barcelona of our paper ‘Pancreatic Islet Isolation With UW Solution: A New Concept’ we had been using collagenase in UW solution in this first short (n=4) series of experiments.
Please note, that I changed the description of the collagenase solution to Hanks’ solution (HBBS) when editing the author proof edition shortly before publication of the final paper August 1990 (PMID_2202137 ; 14) — to best convey our current insights and practice.
For hypothetical reasons only — because the UWS was expected to be detrimental perhaps to cell viablity at the higher temperatures during collagenase digestion of the pancreas (the first stage of islet isolation) — I decided to change the collagenase solution in June 1989 when I began a new series of islet isolations for our first canine islet transplants. From that point on, a modified Hanks’ solution was used for collagenase (17).
We did not observe a difference at the time , whether using one or the other solution for collagenase digestion , with respect to either islet purity, recovery, or viability at islet isolation and days later in culture.
Years later , again tor hypothetical reasons , collagenase in UWS was chosen for the isolation of pig islets, in experiments with a long cold ischemia time of the pancreas during transport with distant pancreas procurement from sows in a slaugtherhouse — Ballering (distant family) in Son — near the south border of The Netherlands, with injection of cold collagenase solution before transport, with the aim of better preservation of the islets (18).
Notes
17) Michael P.M. van der Burg. Pancreatic islet transplantation: studies on the technique and efficacy of islet isolation and transplantation. (Doctoral Thesis, Faculty of Medicine, Leiden University Medical Center (LUMC) , Leiden University), Boskoop: M.P.M. van der Burg | Miracles.Media (ISBN electronic, pdf, 9789080216402 ; ISBN print 9789080216419, 9080216410), 1994: 192 p. Leiden University https://catalogue.leidenuniv.nl/permalink/31UKB_LEU/18s393l/alma9940161419602711
18) Van der Burg MPM, Graham JM. Iodixanol density gradient preparation in University of Wisconsin solution for porcine islet purification. ScientificWorldJournal. 2003 Dec 1;3:1154-9. doi: 10.1100/tsw.2003.107. PMID: 14646009; PMCID: PMC5974767.
Citation info : Author Proof Edition • Michel van der Burg • Miracles.Media • @1MEMO 20241228 • URL michelvanderburg.com/2024/12/28/
Pancreatic Islet Isolation With UW Solution • A New Concept • 20241224
Michel van der Burg • 20241224 • Last update 20241228-1614
In November 1989 in Barcelona, I had my first opportunity to present a paper at a major international organ transplantation meeting — the 4th ESOT — focusing on the rationale for using the UW organ preservation solution during pancreatic islet isolation.
Poster lost
Excited about giving a talk and poster presentation at this 4th ESOT congress in Barcelona, I forgot to take my poster from the overhead compartment of the plane upon arrival in Barcelona. A new poster was made on the spot at the Sandoz booth in the congress venue, by cut and paste work using the manuscript of the congress proceedings paper (PMID 2202137) and enlargement with a Sandoz A3 photocopier. Due to design constraints, the author affiliation info was omitted, and the Du Pont logo was added (image 20241025_1).
Scan of the slide picture taken at the 4th Congress of the European Society for Organ Transplantation, Barcelona (Spain) November 1–4, 1989 • Michel van der Burg • Miracles.Media • 20241025_1 • TakeNode 054301b6-0b6e-458c-84f6-0b76e25092d5
Barcelona ’89 poster remastered
That photo of the poster board in Barcelona (20241025_1) was used for creating this digitally remastered poster presented here (20241025_2) — which now includes the complete manuscript, as well as the Du Pont logo ;)
Poster 4th Congress of the European Society for Organ Transplantation, Barcelona (Spain) November 1–4, 1989 • Digitally remastered from original manuscript of article PMID 2202137 by author, publisher, M.P.M. van der Burg • Miracles.Media • 20241025_2 • TakeNode 124e20db-758a-4bdb-b854-c6cefdb7cc3c
The Nov ’89 Barcelona publications, were the first ones dedicated to presenting the concept , the rationale, for using the UW organ preservation solution during pancreatic islet isolation.
Venue 4th Congress of the European Society for Organ Transplantation, Barcelona (Spain) November 1–4, 1989 • Michel van der Burg • Miracles.Media • 20241224 • TakeNode 103c4c3c-5d36-4111-a5f8-f60e81c5db4d
This wasn’t the first time we reported the news of our results with this new, innovative, method developed in our pre-clinical model for the isolation and purification of pancreatic islets of Langerhans for transplantation in diabetic patients. Below that background, publications, and papers available for download.
Background
I was well aware of the development in the late 80s by Belzer’s team at the University of Wisconsin in Madison (USA) of the new solution to preserve pancreas donor organs during cold storage for transplantation :
In 1986 the Belzer team had reported on the newly developed cold storage solution for pancreas preservation, the University of Wisconsin (UW) Solution (1).
From 1986 our colleague Rutger Ploeg (from the Department of Surgery of Leiden University) had been doing PhD research in Belzer’s organ preservation laboratory in Madison. He introduced the ‘…new cold storage solution, the “UW-Solution” in a letter Dec. 10th, 1987 to all Eurotransplant centres (2), and I had assisted during Rutger Ploeg’s preservation research in Madison on the UW solution in pancreas transplantation in dogs, by calculating the k-values of glucose tolerance (3,4).
1) Wahlberg JA, Southard JH, Belzer FO. Development of a cold storage solution for pancreas preservation. Cryobiology. 1986;23(6):477–482.
2) Alexandre G, Margreiter R, Persijn GG, Pichlmayr R, Terpstra JL, Ploeg RJ. To all Eurotransplant centres : Introduction new cold storage solution, the “UW-Solution”. Letter Dec 10, 1987 , Ploeg RJ, Department of Surgery, University Hospital Leiden (AZL , now LUMC).
3) Ploeg RJ, Goossens D, Sollinger HW, Southard JH, Belzer FO. Efficacy of 48-hour pancreas preservation with UW solution in the dog allograft model. Transplant Proc. 1988 Oct;20(5):1026-8. PMID: 3055478.
4) Ploeg RJ. Preservation of kidney and pancreas with the UW solution : experimental and clinical studies. (Thesis, Leiden University), ‘s-Gravenhage : Pasmans (ISBN 9090042490), 1991: 231 p. URL https://search.worldcat.org/en/title/65672513
First thought
In early 1989, I wondered how our research, into improving the technique of pancreatic islet isolation, could benefit from the great interest in this new organ preservation solution, and the contacts through Ploeg with both the Belzer laboratory and Du Pont (Du Pont Critical Care, Waukegan, USA), the manufacturer of the commercial UW Solution.
Radical choice
The most logical , traditional, choice would be, to investigate organ preservation of the pancreas prior to pancreatic islet isolation. However, logistically, this seemed to be too much of a change in our research. Therefore, in February 1989, I made the radical choice of testing the UW Solution as a replacement for conventional isolation solutions in our preclinical transplantation model of canine islet isolation.
Start
A research plan was drawn up for Du Pont (Robert Carter, Clinical Research Project Coordinator, DuPont, UK), and we started our work March 1989. At first — the first two islet isolation experiments — the UW Solution was only used at the first stage of the isolation procedure, as the collagenase solution for pancreatic injection, transport, and digestion in the laboratory. From April 1989 on, the UW-Solution was used in all our isolation steps, both before density gradient purification, and also for washing the purified islets.
Rationale
The research started in April 1989 with the rationale , that : because the UW-Solution (UWS) protects the viability of the pancreas during cold storage, the UWS can also improve the yield of viable isolated islets, because islet isolation also takes place mainly at low temperature.
Discovery
However, from the first time in April 1989 that the UW-Solution was used during all isolation steps, the tissue immediately appeared different, with almost complete islet purification due to a clear difference in islet density and the darker and more compact looking exocrine pancreatic tissue, compared to the previous conventional isolations.
I did not expect this… but I understood it immediately, even during that first isolation: the effect of UWS during the isolation, the effect on the density, by counteracting cell swelling of especially the other, exocrine, pancreatic tissue, resulting in that tremendous improvement in the purification of islets.
News
The news of the exciting finding of over 90% purity of these UWS-isolated islets was brought first that month, April 1989, during my oral presentation (5) at the General Surgery Science Days, the yearly meeting on the current scientific research taking place at the Department of Surgery of the Leiden University Hospital (now LUMC) in Leiden , Holland .
5) Van der Burg MPM. New methods of islet isolation – a comparative study (dutch : ‘Nieuwe methoden van eilandisolatie – een vergelijkend onderzoek). General Surgery Science Days 1989, Leiden University Hospital (dutch : Wetenschapsdagen Algemene Heelkunde 1989, AZL), Leiden, (The Netherlands) April 28, 1989. Yearly meeting on current research at the Department of Surgery of the Leiden University Hospital (now LUMC) in Leiden , Holland .
Next, a manuscript was submitted June or July 1989 for publication of a Letter in The Lancet (6)… but was rejected.
6) Van der Burg MPM, Gooszen HG, Ploeg RJ, Guicherit OR, Scherft JP, Terpstra JL, Bruijn JA, Frölich M. Pancreatic islet isolation with UW solution: a new concept. Manuscript submitted June/July 1989 for publication as Letter in The Lancet. (rejected). Report online in : Lancet Letter 1989 UW Solution Islet Isolation | 20220823 | Michel van der Burg | michelvanderburg•com
Islet transplantation
Internationally , the news of both our first pre-clinical islet transpant, and the islet purification success when using UWS during islet isolation, was first reported July 8, 1989 at the Anglo Danish Dutch Diabetes Group meeting in Oxford, UK (7). National news reports appeared first September ’89 in the Cicero magazine (8) , and Oct ’89 in the Dutch newspaper Leidsch Dagblad (9).
7). Van der Burg MPM, Gooszen HG, Scherft JP, Terpstra JL, Frölich M, Guicherit OR, Lemkes HHPJ, Bruijn JA. The isolation of canine pancreatic islets for transplantation. Seventh Annual Meeting Anglo Danish Dutch Diabetes Group, Oxford (UK) July 5–9, 1989. Report online in : ADD Diabetes Group 1989, St Edmund Hall, Oxford • Michel van der Burg • Miracles•Media • 20241019 • michelvanderburg•com
8), ‘Eilandjes’ getransplanteerd | by Sylvia van Leeuwen in Cicero, Sep. 15th, 1989 (ISSN 0920-2900), the biweekly publication of the Academic Hospital and Leiden Medical Faculty | 20240830_3 • michelvanderburg•com Report online : Islet Transplantation Breakthrough in Leiden University Hospital • 20240830 • Michel van der Burg • michelvanderburg•com
9). Doorbraak in onderzoek naar behandeling suikerpatiënten. Ontdekking door artsen Leids Academisch Ziekenhuis. (EN tr. “Breakthrough in research into treatment of diabetics. Discovery by doctors of Leiden University Hospital”) | by Gert Visser, in Leidsch Dagblad, Oct 24, 1989 | Dutch newspaper. Report online : Islet Transplantation Breakthrough in Leiden University Hospital • 20240830 • Michel van der Burg • michelvanderburg•com
Internationally at a major meeting, the UWS news was first presented in a poster presentation September 1989 in Minneapolis (10). The UWS data had been added ad hoc in the poster – that is , it wasn’t part of the accepted abstract. The first proceedings paper on our UWS work was also submitted at that meeting , and was published April ’90 (11).
10). Van der Burg MPM, Gooszen HG, Ploeg RJ, Scherft JP, Field MJ, Guicherit OR, Terpstra JL, Van de Woude FT, Frölich M, Bruijn JA. Comparison of islet isolation techniques in dogs: Over 90% purified islets using UW solution. 4th International Symposium on Organ Procurement and Preservation and 2nd International Congress on Pancreatic and Islet Transplantation, Minneapolis (USA) September 17–22, 1989 Report online in : Minneapolis 1989 Poster | 20240804 | Michel van der Burg | michelvanderburg•com
11). Van der Burg MPM, Gooszen HG, Ploeg RJ, Scherft JP, Field MJ, Guicherit OR, Terpstra JL, Van de Woude FJ, Frölich M, Bruijn JA. Comparison of islet isolation techniques in dogs: Over 90% purified islets using UW solution. Transplant Proc. 1990 Apr;22(2):795-6. PMID: 2158174. This paper appears to be not readily available online – a reprint copy is available here PMID_2158174 🔗
A new concept – Nov 1989 , Barcelona, Spain – 4th ESOT
Finally , November 1989 in Barcelona, Spain – at the 4th ESOT meeting – the first presentations followed, focussing on the concept — the rationale — for using the UW organ preservation solution during pancreatic islet isolation — in the book of abstracts (12), the poster shown above (13) that was created from the proceedings paper submitted at this meeting (14), and , I included this UWS work also ad hoc in the talk delivered at this meeting on comparison of islet isolation techniques (12, 15, 16). I plan to post on that talk soon.
As reported in our poster and the manuscript delivered in Barcelona of our paper ‘Pancreatic Islet Isolation With UW Solution: A New Concept’ we had been using collagenase in UW solution in this first short (n=4) series of experiments.
Please note, that I changed the description of the collagenase solution to Hanks’ solution (HBBS) when editing the author proof edition shortly before publication of the final paper August 1990 (PMID_2202137 ; 14) — to best convey our current insights and practice.
For hypothetical reasons only — because the UWS was expected to be detrimental perhaps to cell viablity at the higher temperatures during collagenase digestion of the pancreas (the first stage of islet isolation) — I decided to change the collagenase solution in June 1989 when I began a new series of islet isolations for our first canine islet transplants. From that point on, a modified Hanks’ solution was used for collagenase (17).
We did not observe a difference at the time* , whether using one or the other solution for collagenase digestion , with respect to either islet purity, recovery, or viability at islet isolation and days later in culture.
Years later , collagenase in UWS was chosen for the isolation of pig islets, in experiments with a long cold ischemia time of the pancreas during transport with distant pancreas procurement from sows in a slaugtherhouse — Ballering (distant family) in Son — near the south border of The Netherlands, with injection of cold collagenase solution before transport, with the aim of better preservation of the islets (18).
12) Excerpt from book of abstracts 4th Congress of the European Society for Organ Transplantation, Barcelona (Spain) November 1–4, 1989 , including the abstracts : – Van der Burg MPM, Gooszen HG, Guicherit OR, Ploeg RJ, Scherft JP, Terpstra JL, Bruijn JA, Frölich M. UW solution as the isolation medium markedly improves canine islet isolation (abstract). – Van der Burg MPM, Gooszen HG, Field MJ, Scherft JP, Terpstra JL, Van de Woude FT, Guicherit OR, Frölich M, Bruijn JA. Comparison of current islet isolation techniques in dogs (abstract). Excerpt ESOT abstracts book available here 19891101_ESOT 🔗
13) Van der Burg MPM, Gooszen HG, Ploeg RJ, Guicherit OR, Scherft JP, Terpstra JL, Bruijn JA, Frölich M. Pancreatic islet isolation with UW solution: a new concept. 4th Congress of the European Society for Organ Transplantation, Barcelona (Spain) November 1–4, 1989. Poster presentation. Report online (here) : Pancreatic Islet Isolation With UW Solution – A New Concept • 20241224 • Michel van der Burg • michelvanderburg•com PDF poster — M.P.M. van der Burg • Miracles.Media • 20241025_2 • TakeNode 661fade9-51a7-42b1-a7e1-1e281f74ad6d — available here 20241025_2 🔗
14) Van der Burg MPM, Gooszen HG, Ploeg RJ, Guicherit OR, Scherft JP, Terpstra JL, Bruijn JA, Frölich M. Pancreatic islet isolation with UW solution: a new concept. Transplant Proc. 1990 Aug;22(4):2050-1. PMID: 2202137. This paper appears to be not readily available online – a reprint copy is available here PMID_2202137 🔗
15) Van der Burg MPM, Gooszen HG, Field MJ, Scherft JP, Terpstra JL, Van de Woude FT, Guicherit OR, Frölich M, Bruijn JA. Comparison of current islet isolation techniques in dogs. 4th Congress of the European Society for Organ Transplantation, Barcelona (Spain) November 1–4, 1989. Oral presentation.
16) Van der Burg MPM, Gooszen HG, Field MJ, Scherft JP, Terpstra JL, van de Woude FJ, Guicherit OR, Frölich M, Bruijn JA. Comparison of current islet isolation techniques in dogs. Transplant Proc. 1990 Aug;22(4):2044-5. PMID: 1697121. This paper appears to be not readily available online – a reprint copy is available here PMID_1697121 🔗
17) Michael P.M. van der Burg. Pancreatic islet transplantation: studies on the technique and efficacy of islet isolation and transplantation. (Doctoral Thesis, Faculty of Medicine, Leiden University Medical Center (LUMC) , Leiden University), Boskoop: M.P.M. van der Burg | Miracles.Media (ISBN electronic, pdf, 9789080216402 ; ISBN print 9789080216419, 9080216410), 1994: 192 p. Leiden University https://catalogue.leidenuniv.nl/permalink/31UKB_LEU/18s393l/alma9940161419602711
18) Van der Burg MPM, Graham JM. Iodixanol density gradient preparation in University of Wisconsin solution for porcine islet purification. ScientificWorldJournal. 2003 Dec 1;3:1154-9. doi: 10.1100/tsw.2003.107. PMID: 14646009; PMCID: PMC5974767.
*Audio of data discussion posted in : Percoll UWS Purification Pancreatic Islets • ESOT 1991 Talk • Michel van der Burg • Miracles.Media • @1MEMO 20250114 • URL michelvanderburg.com/2025/01/14/
Updates
20241227-1329 Better copy of Minneapolis proceedings paper replaces PMID_2158174
20241228-1614 Author Proof section on the collagenase solution changes
*20250124-0244 Added reference to audio of data discussion posted in : Percoll UWS Purification Pancreatic Islets • ESOT 1991 Talk • Michel van der Burg • Miracles.Media • @1MEMO 20250114 • URL michelvanderburg.com/2025/01/14/
Citation info : Pancreatic Islet Isolation With UW Solution • A New Concept • Michel van der Burg • Miracles.Media • @1MEMO 20241224 • URL michelvanderburg.com/2024/12/24/
Pancreatic Islet Transplantation • Isolation & Transplantation of Islets of Langerhans • 1988 to early 1990s • Islet Transplant Laboratory • Leiden University Hospital • 20240912_1 (Note 1)
Grants
In 1988, we started a project with unique integration of two important trends in research into the treatment of diabetes by transplantation at the Leiden University Hospital (AZL) in Leiden, with major grants from the dutch Ministry of Welfare, Health and Culture (WVC) and the dutch Diabetes Fund (Diabetes Fonds Nederland).
Grants diabetes research • Newspaper Leidsch Dagblad, June 24, 1988 (Note 2)
The generous grants were received — as reported June 24, 1988 in both the dutch newspaper Leidsch Dagblad (2) and in Cicero, the biweekly publication of the Leiden Medical Faculty and University Hospital (3), as a result of our preliminary research over the previous five years into improving the technique of pancreas transplantation, and more recently the development of a technique for isolating the Islets of Langerhans from the pancreas, which produce hormones such as insulin. The idea behind this is, that these isolated islets, after injection into the body, will restart and maintain insulin production.
Questions
What is the consequence of the loss of normal nerve connections to the islets during transplantation? What are the consequences of transplantation of only a segment of the pancreas, or islet transplantation, when fewer islets are available? What is the consequence after transplantation of the different insulin drainage route, where the insulin does not follow the normal path directly to the liver? To what extent do transplantation procedures disrupt the normal architecture and interactions of islets, the pancreas, and other parts of the gastro-intestinal system, and the blood sugar regulation in the body?
Project
With the hope of answering these questions, the grants supported an ambitious new project supervised by Hein Gooszen, integrating two concomitant PhD projects for detailed islet function studies in both experimental pancreas transplantation, by Onno Guicherit (4) , as well as islet isolation by Michel van der Burg (5) in the same model.
A special feature of the experimental design in this preclinical model is that the results of functional studies in the experimental animal can be compared with such functional studies with the isolated islets from the pancreas, where the direct effect on the islets is investigated during perifusion tests in our laboratory (6).
That same month, June 1988, we began research to improve the method of islet isolation in our laboratory, with the assistance of Jane Field, who had come over that month at our invitation from Minneapolis, where she was a key member of the Surgical Research lab, University of Minnesota (led by David Sutherland).
The introduction of this Minneapolis method for islet isolation significantly improved the yield of isolated islets in our Islet Transplant Lab.
New concept : University of Wisconsin solution (UWS) for islet isolation
Next , early 1989 , our Islet Transplant Lab took a major step forward in a new approach to isolation and purification of the islets by replacing the islet isolation solution (the general used basic physiological salt solution or tissue culture solution) with a novel organ preservation solution : the University of Wisconsin solution (UWS). The method of choice nowadays in clinical islet transplantation centers. More on this introduction of UWS will soon be posted here. A reprint of a first poster presented in Minneapolis, Sep. 1989, is now online here (7).
Program
June 1989 our Surgery Department (Leiden University) started an additional project, for ‘auto’ transplantation of isolated islets : Isolation of the islets from the canine pancreas and transplantation of the isolated islets ca 4 hours later in the same animal (8).
Thus, from 1989, we studied the insulin secretion of the isolated islets in perifusion experiments in the laboratory (‘in vitro’) in comparison with both the islet function ‘in vivo’ after transplantation of the isolated islets, and in the concomitant pancreas transplantation experiments — in support of our clinical pancreas transplantation program in Leiden.
Notes
1) Pancreatic Islet Transplantation • Isolation & Transplantation of Islets of Langerhans • 1988 to early 1990s • Islet Transplant Laboratory • Leiden University Hospital • 20240912_1 • michelvanderburg•com | TakeNode 5b5ca362-2adf-497a-b2be-a83e47617017 | Image edit based on : Pancreatic islet transplantation • Michel van der Burg • Miracles.Media • Thesis Repository Leiden University https://hdl.handle.net/1887/3486604
Captions in Note 9
2) Subsidie voor onderzoek suikerziekte. Leidsch Dagblad, June 24, 1988 (leiden.courant.nu) • 20240912_2 • michelvanderburg•com
3) Diabetesonderzoek | AZL krijgt grote subsidies voor diabetesonderzoek | by Sylvia van Leeuwen in Cicero, June 24th, 1988 (ISSN 0920-2900), the biweekly publication of the Academic Hospital and Leiden Medical Faculty | 20240912_3 • michelvanderburg•com | Download PDF article below : file 20240912_3_CICERO
4) Onno R. Guicherit. Long-term metabolic sequelae of beta cell mass reduction, systemic venous drainage and denervation of the canine pancreas : experimental studies in relation to clinical pancreas transplantation. (Doctoral Thesis, Faculty of Medicine, Leiden University Medical Center (LUMC) , Leiden University)(S.l.: s.n.), (ISBN print: 9789090072319, 9090072314), 1994: 149 p. URL WorldCat https://search.worldcat.org/en/title/69433794
5) Michael P.M. van der Burg. Pancreatic islet transplantation: studies on the technique and efficacy of islet isolation and transplantation. (Doctoral Thesis, Faculty of Medicine, Leiden University Medical Center (LUMC) , Leiden University), Boskoop: M.P.M. van der Burg | Miracles.Media (ISBN electronic, pdf, 9789080216402 ; ISBN print 9789080216419, 9080216410), 1994: 192 p. URL Thesis Repository Leiden University https://hdl.handle.net/1887/3486604
9) Pancreatic Islet Transplantation • Isolation & Transplantation of Islets of Langerhans 1988 to early 1990s • Islet Transplant Laboratory • Leiden University Hospital • 20240912_1 • michelvanderburg•com | TakeNode 5b5ca362-2adf-497a-b2be-a83e47617017 |
Captions Image 20240912_1 :
1. Paul Langerhans
2. Section of the normal dog pancreas showing islets stained reddish-brown (immunostained for insulin)
3. Michel van der Burg, watching the dog islet isolation procedure in the Minneapolis surgical research laboratory (University of Minnesota), September 1989, with Philippe Morel and Pericles Tzardis
4. Experimental Surgery Laboratory in Leiden, June 1988, with (left to right) Hein Gooszen, Jane Field (Minneapolis), and Onno Guicherit, starting the surgical procedure for canine islet isolation
5. The pancreas (segment) is removed
6. Islet Transplant Laboratory in Leiden, at the start of islet isolation, with infusion of the collagenase solution via the ducts in the dog pancreas (whole gland for islet transplantation) – collagenase leaking from the pancreas is recirculated using a roller pump
7. During collagenase digestion at 37-39°C the pancreas falls apart, shown here in a low magnification microscopy image of pancreatic exocrine tissue, and a free-ed small blood vessel
8. Dissociated pancreatic tissue, on ice
9. Tissue is further dispersed in the cold isolation solution (here RPMI tissue culture solution is used), by aspiration in a syringe, and sieved to remove undigested fragments, ducts and vessels ~ demonstrated here by Jane Field (Minneapolis), with the introduction of the Minneapolis isolation procedure, June 1988, in our Islet Transplant Laboratory in Leiden
10. Microscopy of tissue suspension, with a low purity of the islets (stained red by dithizone) obtained by density separation in Dextran gradients (in Hanks’ solution) after islet isolation in the RPMI tissue culture solution (dark-brown exocrine fragments remain unstained)
11. Pure islet suspension obtained by density separation in Percoll gradients (in University of Wisconsin solution) after islet isolation in the University of Wisconsin solution (islets are only slightly stained due to poor diffusion over the cell membrane in the preservation solution)
12. Section of highly purified islets obtained by Dextran density separation after isolation in the University of Wisconsin solution
13. Islets are autotransplanted by infusion in the spleen of the dog
14. Transplanted islet in section of the spleen of one of the dogs shortly after the onset of fasting hyperglycemia at three months posttransplant (immunostained for insulin), 1989
15. Highly purified human islets obtained by Percoll density separation after isolation in the University of Wisconsin solution (immunostained with gold for insulin; not counterstained), July 1990
Pancreatic islet transplantation • Michel van der Burg • Miracles.Media | QR code (link to Thesis Repository Leiden University) https://hdl.handle.net/1887/3486604
Citation info : Pancreas & Islet Transplantation Program • 20240912 | Michel van der Burg | michelvanderburg•com
“Breakthrough in research into treatment of diabetics. Discovery by doctors of Leiden University Hospital” …headlines the Dutch newspaper ‘Leidsch Dagblad’ (1).
The dutch newspaper ‘Leidsch Dagblad’ reports October 24, 1989, on a breakthrough in the treatment of diabetic patients…successful transplantation of the insulin producing ‘islets’ in dogs, after isolation of the islets using the UW organ storage solution (UWS)
The first successful transplant June 13, 1989 was a big step. Generally multiple pancreases are needed for an adequate number of purified islets for one transplant. Here the dog’s pancreas was used for the isolation of islets, that were injected back in the animals’ spleen.
‘The breakthrough in the research came when Van der Burg came up with the idea of using the UW organ storage solution — developed several years ago at the University of Wisconsin — for the preservation of the islets. This turned out to yield good results. The yield of pure islets could be increased using this method.’
Cicero
News based on story published Sep. 15th, 1989 by journalist Sylvia Van Leeuwen in Cicero, the biweekly publication of the Academic Hospital and Leiden Medical Faculty (2).
Ploeg, Gooszen and Van der Burg (left to right) discuss the achieved results, Monday Sep 4, 1989 at the Department of Surgery, University Hospital Leiden. Photo Tejo Ringers • 20240830_2 • michelvanderburg•com • TakeNode 3fc69a06-04bf-4f0d-bc77-cedb6d7b7f75
One more thing
We reported long-term function of the islet transplants (3,4). Failure after 3.5 years of a well-functioning islet transplant is puzzling. Nowadays, in clinical islet transplantation, failure of long-term functioning grafts in diabetic patients is still puzzling. Therefore, note (data not published before), that body weight of this autografted animal had increased to almost 150%, from 12.5 kg at the time of islet transplantation, to 18.0 kg at the time of graft failure 3.5 years later (Dec. 1992) — obviously, with an increased insulin demand that may have contributed to graft failure.
Notes
1) Doorbraak in onderzoek naar behandeling suikerpatiënten. Ontdekking door artsen Leids Academisch Ziekenhuis. (EN tr. “Breakthrough in research into treatment of diabetics. Discovery by doctors of Leiden University Hospital”) | by Gert Visser, in Leidsch Dagblad, Oct 24, 1989 | Dutch newspaper, URL https://leiden.courant.nu/issue/LD/1989-10-24/edition/0/page/15
The background in the image here (20240830_1), is the original color slide of the ‘Islets of Langerhans’ showing in the newspaper.
2) ‘Eilandjes’ getransplanteerd | by Sylvia van Leeuwen in Cicero, Sep. 15th, 1989 (ISSN 0920-2900), the biweekly publication of the Academic Hospital and Leiden Medical Faculty | 20240830_3 • michelvanderburg•com Download PDF article, file 20240830_3_CICERO
4) Van der Burg MPM, Guicherit OR, Jansen JBMJ, Frölich M, Lamers CBHW, Lemkes HHPJ, Bruijn JA, Gooszen HG. Function and survival of intrasplenic islet autografts in dogs. Diabetologia 39, 37–44 (1996). https://doi.org/10.1007/BF00400411
5) Van der Burg MPM, Gooszen HG, Ploeg RJ, Guicherit OR, Scherft JP, Terpstra JL, Bruijn JA, Frölich M. Pancreatic islet isolation with UW solution: a new concept. 4th Congress of the European Society for Organ Transplantation, Barcelona (Spain) November 1–4, 1989.
6) The islet transplant news was first reported by me at an invited lecture July 8, 1989 for the Anglo Danish Dutch Diabetes Group meeting in St Edmund Hall, University of Oxford, Oxford (UK) : Van der Burg MPM, Gooszen HG, Scherft JP, Terpstra JL, Frölich M, Guicherit OR, Lemkes HHPJ, Bruijn JA. The isolation of canine pancreatic islets for transplantation. Seventh Annual Meeting Anglo Danish Dutch Diabetes Group, Oxford (UK) July 5–9, 1989.
7) Leiden Islet Laboratory History • 20240803 | Michel van der Burg | michelvanderburg•com
Citation info : Islet Transplantation Breakthrough in Leiden University Hospital • 20240830 • Michel van der Burg • michelvanderburg•com
Isolation of Islets of Langerhans from the Canine Pancreas | Fall meeting of the Dutch Association for Calcium and Bone Metabolism, Leiden (The Netherlands) November 13, 1987 • 20240829_1 • michelvanderburg•com • TakeNode 6b80e3a7-af16-41f7-b15f-cdeb85e377f3
Cradle of Islet Transplants
The Laboratory of Cell Biology and Histology in Leiden (headed by Prof J.P. Scherft), was the cradle of our Islet Transplantation Lab, where we made a fresh start Summer 1986 (1), and presented our first poster the following year.
Poster
Our first year of research was presented in this poster (Image 20240829_1) at the Fall Meeting Friday, November 13, 1987 , of the Dutch Association for Calcium- and Bone-metabolism*, in Leiden, Holland (2) in that Cell Biology building of the University Hospital Leiden (‘Building 55’) where our Islet Lab was hosted for the first decade.
The poster shows the results of islet isolations from canine pancreas segments performed with a method developed for human islet isolation by the surgeon Derek Gray (Oxford, UK) with some modifications (3). Pancreas segments were harvested at surgery for our concommitant research on segmental pancreas transplantation in this pre-clinical model (4,5,6,7).
Isolation of Islets of Langerhans from the Canine Pancreas | Islet Transplantation Laboratory, Leiden University Hospital, 1987 • 20240829_2 • michelvanderburg•com • TakeNode a9a389d9-c06f-467a-9ee0-1c2dc5f2403a
The photo collage (Image 20240829_2) also shown in the Methods section of the poster illustrates, resp. : 1) removal of a canine pancreatic segment at surgery; 2) distension of the gland by ductal injection of a collagenase solution; 3) the mucoid appearance after collagenase digestion of the connective tissue in the gland for 20-25 min at 39°C; 4) the digestion end-point, when gentle contusion of a sample of the tissue appears to liberate free islets from the surrounding exocrine tissue; 5) dispersion of the tissue with forceps and aspiration through needles; 6) microscopy of the suspension of purified islets (arrow denotes islet) contaminated with some exocine fragments , after sieving and Ficoll density gradient centrifugation to separate the islets from most of the exocrine fragments.
Figure 1 of the Results shows the relation between islet size and number of the isolated islets. Quantitation of the total volume of islets in the pancreas, and after islet isolation , demonstrated that on average 5-10% of the islets in the pancreas are isolated after purification. Figure 2 shows the timeline of insulin secretion by the isolated islets, in response to stimulation with glucose (sugar) during perifusion with a basic salts solution containing low -, high -, and low sugar levels, resp.
The posters were produced at the Cell Biology department using a pen plotter machine. A corridor in the building with part of the gallery of posters was photographed at the meeting (Image 20240829_3).
Cell Biology – 40th anniversary, 1947-1987
Laboratory of Cell Biology and Histology, Leiden (Holland) celebrating the 40th anniversary, 1947-1987. Front entrance ‘Building 55’ University Hospital Leiden, November 14, 1987 20240829_4 • michelvanderburg•com • TakeNode bcf4674c-0cc7-4545-96b1-1a8e0c451150
The 40th anniversary of the Cell Biology Department was celebrated 13-14 November 1987 (with a banner and flags at the entrance of the building (Image 20240829_4) , with the science meeting Nov. 13th and a get-together with (former) co-workers, including Professor emiritus Pieter J. Gaillard, dutch pioneer of tissue culture and transplantation, and founder of the laboratory when appointed professor of Experimental Histology at the Leiden University, Friday, November 14, 1947 (8,9).
Professor Gaillard’s lab started in 1947 in the cellar of the Anatomical Laboratory, and moved in 1953-1954 to ‘Building 55’ of the University Hospital Leiden; shown above. The name was changed to Cell Biology and Histology, in 1964. Cell Biology is currently part of the Department Of Cell & Chemical Biology , and housed within the research building of the Leiden University Medical Centre.
I photographed the emiritus Gaillard receiving a ‘souvenir’ from Professor Scherft (Image 20240829_5) at the 40th anniversary of the Cell Biology Laboratory in Leiden (Holland), Nov. 14, 1987.
P.J. Gaillard (on the left) & J.P. Scherft, Cell Biology Laboratory in Leiden (Holland), Nov. 14, 1987 • 20240829_5 • michelvanderburg•com • TakeNode 08054d8f-c978-422f-9f3f-5c8507cf8869
First clinical islet cell transplant
In 1944, during World War II, Gaillard and the Leiden surgeon P.J. Kooreman performed the first successfull clinical (allogeneic) transplant of (cultured) islet cells in Leiden. Dutch newspapers reported (10), Gaillard had spoken at a meeting in 1949 organized by local hospitals in The Hague, Holland, on some of his early transplants in diabetic patients, with success in one patient during the war (Image 20240829_6) : “Prof. Gaillard considers it too early to say much about the possibilities of this intervention. Although he could report that a female patient can be considered cured in nine months. A bombing took her life, so that further observation was cut off.” . The Islet Transplant Registry listed the transplant as the 5th performed in the world (6). Gaillard & Kooreman also performed a successful series of transplants of cultured parathyroid tissue , a firm base of what would become the focus of Gaillards research and the Cell Biology Laboratory , the hormonal regulation of bone metabolism (9,10,11,12).
Prof. Dr. P.J. Gaillard spoke about successes in transplant trials. Nieuwe Leidsche Courant, Oct. 1, 1949 • 20240829_6 • michelvanderburg•com • TakeNode c5be6800-f3da-453e-9d40-d8fafffd27aa
Introduction of collagenase isolation of islets
Also, of note, is the first introduction of pancreatic islet isolation by collagenase digestion in 1965 by the polish researcher Stanislaw Moskalewski with the help of Gaillard working at the Cell Biology Laboratory (13) — the basis of large scale islet isolation and transplantation.
Notes
1) Leiden Islet Laboratory History • 20240803 | Michel van der Burg | michelvanderburg•com
2) Van der Burg MPM, Gooszen HG, Terpstra JL, Scherft JP, Frölich M, Lemkes HHPJ. Isolation of the islets of Langerhans from the canine pancreas. Fall meeting of the Dutch Association for Calcium and Bone Metabolism, Leiden (The Netherlands) November 13, 1987.
3) Gray DWR, McShane P, Grant A, Morris PJ. A Method for Isolation of Islets of Langerhans from the Human Pancreas. Diabetes 1 November 1984; 33 (11): 1055–1061. https://doi.org/10.2337/diab.33.11.1055
4) Gooszen HG. Canine segmental pancreatic autotransplantation. Analysis of the effects of ductobliteration. Thesis, Leiden, 1984
5) Van der Burg MPM, Gooszen HG, Guicherit OR, Jansen JBMJ, Frölich M, Haastert FA, Lamers CBHW. Contribution of partial pancreatectomy, systemic hormone delivery and duct obliteration to glucose regulation in canine pancreas: Importance in pancreas transplantation. Diabetes 1989; 38: 1082–1089.
6) Pancreatic islet transplantation (doctoral thesis, 1994) by Michel van der Burg. ISBN 9789080216402. Scholarly Publications repository of Leiden University. Persistent URL https://hdl.handle.net/1887/3486604
Eerste alvleesklier-transplantatie | Nederlandse primeur in Academisch Ziekenhuis Leiden. Leidsch Dagblad, May 18, 1984 (leiden.courant.nu) • 20240829_7 • michelvanderburg•com • TakeNode 5d3c88a5-4c67-48fd-8a8c-8165a5c2edfe
8) Van CH0 naar CH40 met Ca2+ als katalysator. Scherft J.P. In : Laboratorium voor Celbiologie en Histologie 1947-1987 | Uitgave ter gelegenheid van het veertigjarig bestaan van het Laboratorium voor Celbiologie en Histologie van de Medische Faculteit te Leiden.
9) Enkele notities over het begin van de weefselkweek in Leiden. Gaillard P.J. In : Laboratorium voor Celbiologie en Histologie 1947-1987 | Uitgave ter gelegenheid van het veertigjarig bestaan van het Laboratorium voor Celbiologie en Histologie van de Medische Faculteit te Leiden.
10) Prof. dr P.J. Gaillard sprak over successen bij transplantatie-proeven. Nieuwe Leidsche Courant, Oct. 1, 1949. | Dutch newspaper , URL https://leiden.courant.nu/issue/NLC/1949-10-01/edition/null/page/2 | “Prof. Gaillard acht het nog te vroeg om over de mogelijkheden van deze ingreep veel te zeggen. Al kon hij mededelen, dat een vrouwelijke patiënt in negen maanden tijds als genezen beschouwd kan worden. Een bombardement benam haar het leven, zodat men van verdere observatie werd afgesneden”.
12) Jon van Rood: The pioneer and his personal view on the early developments of HLA and immunogenetics. Martine J. Jagera M.J, Brand A, Claas F.H.J. Transplant Immunology 52 (2019) 1–26
* Strange to find the Dutch Association for Calcium and Bone Metabolism indicates online a start in 1990 – while I visited their fall meeting in 1987 (see also refs 8,9)
Update (20240912) correction text perifusion solution
Citation info : Cradle of Islet Transplants • 20240829 • Michel van der Burg • michelvanderburg•com
