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Evaluation of the new pancreas preservation solution, I-Let Protect, for clinical islet isolation and transplantation
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology. (Olle Korsgren)ORCID iD: 0000-0003-4511-6149#sthash.NKvqlgKU.dpuf
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2015 (English)Manuscript (preprint) (Other academic)
Abstract [en]


This prospective study aimed to evaluate polydimethylsiloxane 5 (F6H8S5) preservation of pancreases in a clinical setting compared with standard solutions for 1) cold ischemia-time (CIT) <10 hour and 2) a prolonged CIT > 20 hour.


Part 1. Procured clinical grade pancreases were shipped in either F6H8S5 or in standard preservation solutions, i.e., University of Wisconsin (UW) or Custodiol. F6H5S5 were pre-oxygenated for at least 15 minutes.

Part 2. Included clinical grade pancreases were procured in UW or Custodiol. Upon arrival at the islet isolation laboratory, duodenum was removed followed by rough trimming while F6H8S5 was oxygenated for 15-20 minutes. Trimmed pancreases were immersed into oxygenated F6H8S5 and stored at 4°C over night followed by subsequent islet isolation.


Pancreas preservation using F6H8S5 proved as effective as UW and Custadiol when used within CIT up to 10 hour, both in terms of isolation outcome and islet functionality. Preservation in F6H8S5 of pancreases with prolonged CIT gave results similar to controls with CIT< 10 hours for both isolated islet functionality and isolation outcome.


This prospective study of clinically obtained pancreases indicate a clear benefit of using F6H8S5 on pancreases with prolonged CIT when compared with other organ preservation solutions. F6H8S5 preserved islet quality and quantity compared with islets isolated from pancreases with CIT of less than 10 hour.

Place, publisher, year, edition, pages
National Category
Endocrinology and Diabetes
URN: urn:nbn:se:uu:diva-254357OAI: oai:DiVA.org:uu-254357DiVA: diva2:843904
Available from: 2015-07-31 Created: 2015-06-08 Last updated: 2015-08-28
In thesis
1. Technical challenges in human islet isolation
Open this publication in new window or tab >>Technical challenges in human islet isolation
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Transplantation of islets of Langerhans is an effective treatment option for patients with brittle type 1 diabetes mellitus. This treatment restores glucose control and also reduces hypoglycemia. Unfortunately, the outcome from islet isolations is variable, and many preparations do not yield sufficient islet number or islet quality.

The aim of this thesis was to improve the isolation procedure, thereby making more preparations available for clinical transplantation.

A well-established method for pathogen inactivation was applied to human serum used in the islet isolation process. Evaluation of isolated islets stored in medium supplemented with pathogen-inactivated serum showed that pathogen inactivation did not have negative effects. These findings will enable the use of human serum in clinical cell transplantation programs, while simultaneously increasing patient safety.

Pre-incubation of islets prior to gradient separation is an established standard in the field of islet isolation. Through a reduction in the pre-incubation step, isolation time could be reduced by almost an hour without affecting the isolation outcome.

A commercially available protease enzyme, clostripain, was added to the enzyme blend used in islet isolation. Addition of clostripain was found to increase the number of islets isolated as well as the purified tissue volume and fulfillment of transplant criteria. Use of clostripain should help to increase the number of successful isolations.

A newly developed pancreas-specific preservation solution, I-Let protect, was evaluated. As compared to standard preservation solutions, it can be used in situations of prolonged cold ischemic time without affecting the isolation outcome or islet functionality. I-Let protect can also be used in establishing a protocol that would eliminate the need for night- time isolations.

Through the work in this thesis, several key elements in human islet isolation have been optimized, and further knowledge has been gained.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2015. 57 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 1119
National Category
Endocrinology and Diabetes
urn:nbn:se:uu:diva-259358 (URN)978-91-554-9283-0 (ISBN)
Public defence
2015-09-18, Auditorium Minus, Gustavianum, Akademigatan 3, Uppsala, 09:15 (English)
Available from: 2015-08-27 Created: 2015-07-31 Last updated: 2015-10-01

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Ståhle, Magnus
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