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Human Islet Isolation Processing Times Shortened By One Hour: Minimized Incubation Time Between Tissue Harvest and Islet Purification
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Clinical Immunology. (Korsgren)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Clinical Immunology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Clinical Immunology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Clinical Immunology. (Korsgren)
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2013 (English)In: Transplantation, ISSN 0041-1337, E-ISSN 1534-6080, Vol. 96, no 12, E91-E93 p.Article in journal, Letter (Refereed) Published
Place, publisher, year, edition, pages
2013. Vol. 96, no 12, E91-E93 p.
National Category
Endocrinology and Diabetes
Identifiers
URN: urn:nbn:se:uu:diva-220216DOI: 10.1097/01.tp.0000437562.31212.d5ISI: 000330439900004PubMedID: 24342944OAI: oai:DiVA.org:uu-220216DiVA: diva2:705620
Available from: 2014-03-17 Created: 2014-03-12 Last updated: 2017-12-05Bibliographically approved
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.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 1119
National Category
Endocrinology and Diabetes
Identifiers
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)
Opponent
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Available from: 2015-08-27 Created: 2015-07-31 Last updated: 2015-10-01

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Ståhle, MagnusKorsgren, Olle

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