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Mammalian Tissue-Free Liberase: A New GMP-Graded Enzyme Blend for Human Islet Isolation
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Oncology, Radiology and Clinical Immunology, Clinical Immunology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Oncology, Radiology and Clinical Immunology, Clinical Immunology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Oncology, Radiology and Clinical Immunology, Clinical Immunology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Oncology, Radiology and Clinical Immunology, Clinical Immunology.
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2010 (English)In: Transplantation, ISSN 0041-1337, E-ISSN 1534-6080, Vol. 90, no 3, 332-333 p.Article in journal, Letter (Refereed) Published
Place, publisher, year, edition, pages
2010. Vol. 90, no 3, 332-333 p.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-135881DOI: 10.1097/TP.0b013e3181e117e3ISI: 000280581200020OAI: oai:DiVA.org:uu-135881DiVA: diva2:376926
Available from: 2010-12-13 Created: 2010-12-09 Last updated: 2017-12-11Bibliographically approved
In thesis
1. Islet Transplantation a Technical Challenge: Studies on Human Pancreas Preservation and Enzymatic Digestion
Open this publication in new window or tab >>Islet Transplantation a Technical Challenge: Studies on Human Pancreas Preservation and Enzymatic Digestion
2011 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Islet transplantation has found its niche in diabetes treatment. It has contributed to a better quality of life and better glycemic control of patients with diabetes suffering from severe hypoglycemia that are not eligible for vascularized pancreas transplantation. Islet isolation is a technically challenging procedure. The different studies within this doctoral thesis aim to improve and standardize different steps in the isolation procedure. They are in particular looking to improve human pancreas preservation during cold storage, to optimize islet release from the exocrine tissue and to assess whether the isolated islet yield can be predicted from a biopsy.

We found that pancreas preservation with pre-oxygenated perfluorodecalin (two-layer method) did not improve the ischemic tolerance of the human pancreas as compared to cold storage with the University of Wisconsin (UW) solution. Furthermore, in pancreas with long cold ischemia time (CIT) (>10 hours), Histidine-Tryptophan-Ketoglutarate (HTK) had a limited preservation capacity as compared with the UW solution with respect to isolation outcome. We also found that during enzymatic pancreas digestion, Vitacyte HA was able to provide a similar islet yield and quality as Serva NB1 with less collagenase activity and shorter digestion time. We further describe the first experience with a new GMP manufactured enzyme called Liberase MTF-S for successful human islet isolation. Finally, we found that the isolated islet yield could not be predicted from a biopsy taken from the head of the pancreas concerning solely morphological parameters of the islets tissue.

The improvement of pancreas preservation will allow for marginal organs with prolonged cold ischemia time to expand the donor pool. Better knowledge of how the pancreatic extracellular matrix is digested by collagenase will lead to a fast and predictable islet release from the exocrine tissue. By standardizing the isolation procedure and improving organ selection we will increase the success rate in human islet isolation, thereby making islet transplantation available for more patients.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2011. 58 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 692
Keyword
Islet transplantation, Human islet isolation, Pancreas preservation, Two-layer method, Perfluorocarbon, Cold storage, Cold ischemia, University of Wisconsin solution, Histidine-tryptophan-ketoglutarate, HTK, Pancreas preservation, Prediction, Collagenase, Trasplante de islotes, Aislamiento de islotes, Preservación del pancreas, Método de las dos capas, Colagenasa, Solución de Wisconsin, HTK, Histidina-triptófano-ketoglutarato, Predicción
National Category
Endocrinology and Diabetes Surgery
Research subject
Endocrinology and Diabetology; Cell Research; Clinical Immunology
Identifiers
urn:nbn:se:uu:diva-156893 (URN)978-91-554-8126-1 (ISBN)
Public defence
2011-09-23, Rudbecksalen, Rudbecklaboratoriet. Dag Hammarskjölds väg 20. SE-751 85, Uppsala, 13:15 (English)
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Supervisors
Available from: 2011-09-02 Created: 2011-08-10 Last updated: 2011-11-03Bibliographically approved

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