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Ståhle, Magnus U.
Alternative names
Publications (10 of 12) Show all publications
Eich, T., Ståhle, M. U., Gustafsson, B., Horneland, R., Lempinen, M., Lundgren, T., . . . Korsgren, O. (2018). Calcium: A Crucial Potentiator for Efficient Enzyme Digestion of the Human Pancreas. Cell Transplantation, 27(7), 1031-1038
Open this publication in new window or tab >>Calcium: A Crucial Potentiator for Efficient Enzyme Digestion of the Human Pancreas
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2018 (English)In: Cell Transplantation, ISSN 0963-6897, E-ISSN 1555-3892, Vol. 27, no 7, p. 1031-1038Article in journal (Refereed) Published
Abstract [en]

Background: Effective digestive enzymes are crucial for successful islet isolation. Supplemental proteases are essential because they synergize with collagenase for effective pancreatic digestion. The activity of these enzymes is critically dependent on the presence of Ca2+ ions at a concentration of 5–10 mM. The present study aimed to determine the Ca2+ concentration during human islet isolation and to ascertain whether the addition of supplementary Ca2+ is required to maintain an optimal Ca2+ concentration during the various phases of the islet isolation process.

Methods: Human islets were isolated according to standard methods and isolation parameters. Islet quality control and the number of isolations fulfilling standard transplantation criteria were evaluated. Ca2+ was determined by using standard clinical chemistry routines. Islet isolation was performed with or without addition of supplementary Ca2+ to reach a Ca2+ of 5 mM.

Results: Ca2+ concentration was markedly reduced in bicarbonate-based buffers, especially if additional bicarbonate was used to adjust the pH as recommended by the Clinical Islet Transplantation Consortium. A major reduction in Ca2+ concentration was also observed during pancreatic enzyme perfusion, digestion, and harvest. Additional Ca2+ supplementation of media used for dissolving the enzymes and during digestion, perfusion, and harvest was necessary in order to obtain the concentration recommended for optimal enzyme activity and efficient liberation of a large number of islets from the human pancreas.

Conclusions: Ca2+ is to a large extent consumed during clinical islet isolation, and in the absence of supplementation, the concentration fell below that recommended for optimal enzyme activity. Ca2+ supplementation of the media used during human pancreas digestion is necessary to maintain the concentration recommended for optimal enzyme activity. Addition of Ca2+ to the enzyme blend has been implemented in the standard isolation protocols in the Nordic Network for Clinical Islet Transplantation.

Keywords
calcium, clinical islet transplantation, diabetes, islet isolation
National Category
Surgery
Identifiers
urn:nbn:se:uu:diva-364508 (URN)10.1177/0963689718779350 (DOI)000440338700003 ()29945463 (PubMedID)
Available from: 2018-11-05 Created: 2018-11-05 Last updated: 2018-11-05Bibliographically approved
Svensson, E., Milenova, I., Wenthe, J., Ståhle, M., Leja-Jarblad, J., Ullenhag, G., . . . Loskog, A. S. (2017). Shaping the Tumor Stroma and Sparking Immune Activation by CD40 and 4-1BB Signaling Induced by an Armed Oncolytic Virus.. Clinical Cancer Research, 23(19), 5846-5857
Open this publication in new window or tab >>Shaping the Tumor Stroma and Sparking Immune Activation by CD40 and 4-1BB Signaling Induced by an Armed Oncolytic Virus.
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2017 (English)In: Clinical Cancer Research, ISSN 1078-0432, E-ISSN 1557-3265, Vol. 23, no 19, p. 5846-5857Article in journal (Refereed) Published
Abstract [en]

Purpose: Pancreatic cancer is a severe indication with short expected survival despite surgery and/or combination chemotherapeutics. Checkpoint blockade antibodies are approved for several cancer indications, but pancreatic cancer has remained refractory. However, there are clinical data suggesting that stimulation of the CD40 pathway may be of interest for these patients. Oncolytic viruses armed with immunostimulatory genes represent an interesting approach. Herein, we present LOAd703, a designed adenovirus armed with trimerized CD40L and 4-1BBL that activates the CD40 and 4-1BB pathways, respectively. As many cells in the tumor stroma, including stellate cells and the infiltrating immune cells, express CD40 and some 4-1BB, we hypothesize that LOAd703 activates immunity and simultaneously modulates the biology of the tumor stroma.Experimental Design: Tumor, stellate, endothelial, and immune cells were infected by LOAd703 and investigated by flow cytometry, proteomics, and functional analyses.Results: LOAd703-infected pancreatic cell lines were killed by oncolysis, and the virus was more effective than standard-of-care gemcitabine. In in vivo xenograft models, LOAd703 efficiently reduced established tumors and could be combined with gemcitabine for additional effect. Infected stellate and tumor cells reduced factors that promote tumor growth (Spp-1, Gal-3, HGF, TGFβ and collagen type I), while chemokines were increased. Molecules involved in lymphocyte migration were upregulated on infected endothelial cells. Dendritic cells were robustly stimulated by LOAd703 to produce costimulators, cytokines and chemokines, and such DCs potently expanded both antigen-specific T cells and NK cells.Conclusions: LOAd703 is a potent immune activator that modulates the stroma to support antitumor responses. Clin Cancer Res; 1-12. ©2017 AACR.

National Category
Cancer and Oncology Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:uu:diva-330158 (URN)10.1158/1078-0432.CCR-17-0285 (DOI)000412160500021 ()28536305 (PubMedID)
Funder
Swedish Cancer Society
Available from: 2017-09-27 Created: 2017-09-27 Last updated: 2018-02-02Bibliographically approved
Ståhle, M., Foss, A., Gustafsson, B., Lempinen, M., Lundgren, T., Rafael, E., . . . Friberg, A. (2016). Evaluation of Perfluorohexyloctane/Polydimethylsiloxane for Pancreas Preservation for Clinical Islet Isolation and Transplantation. Cell Transplantation, 25(12), 2269-2276
Open this publication in new window or tab >>Evaluation of Perfluorohexyloctane/Polydimethylsiloxane for Pancreas Preservation for Clinical Islet Isolation and Transplantation
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2016 (English)In: Cell Transplantation, ISSN 0963-6897, E-ISSN 1555-3892, Vol. 25, no 12, p. 2269-2276Article in journal (Refereed) Published
Abstract [en]

This study aimed to evaluate a 50:50 mix of perfluorohexyloctane/polydimethylsiloxane 5 (F6H8S5) preservation of pancreases in a clinical setting compared with standard solutions for 1) cold ischemia time (CIT) <10 h and 2) an extended CIT >20 h. Procured clinical-grade pancreases were shipped in either F6H8S5 or in standard preservation solutions, that is, University of Wisconsin (UW) or Custodiol. F6H5S5 was preoxygenated for at least 15 min. Included clinical-grade pancreases were procured in UW or Custodiol. Upon arrival at the islet isolation laboratory, the duodenum was removed followed by rough trimming while F6H8S5 was oxygenated for 15-20 min. Trimmed pancreases were immersed into oxygenated F6H8S5 and stored at 4 C overnight followed by subsequent islet isolation. Pancreas preservation using F6H8S5 proved as effective as UW and Custadiol when used within CIT up to 10 h, in terms of both isolation outcome and islet functionality. Preservation in F6H8S5 of pancreases with extended CIT gave results similar to controls with CIT <10 h for both isolated islet functionality and isolation outcome. This study of clinically obtained pancreases indicates a clear benefit of using F6H8S5 on pancreases with extended CIT as it seems to allow extended cold ischemic time without affecting islet function and islet numbers.

Keywords
Islet isolation, Pancreas preservation, Clinical islet transplantation, Perfluorohydrocarbons, Oxygenation, Hypoxia
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-313431 (URN)10.3727/096368916X691709 (DOI)000390183200014 ()
Funder
Swedish Research Council, K2015-54X-12219-19-4Swedish Child Diabetes FoundationSwedish Diabetes AssociationNIH (National Institute of Health), 2U01AI065192-06EXODIAB - Excellence of Diabetes Research in Sweden
Available from: 2017-01-30 Created: 2017-01-19 Last updated: 2017-11-29Bibliographically approved
Ståhle, M., Foss, A., Gustafsson, B., Lempinen, M., Lundgren, T., Rafael, E., . . . Friberg, A. (2015). Clostripain, the Missing Link in the Enzyme Blend for Efficient Human Islet Isolation. Transplantation Direct, 1(5), 1-6
Open this publication in new window or tab >>Clostripain, the Missing Link in the Enzyme Blend for Efficient Human Islet Isolation
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2015 (English)In: Transplantation Direct, ISSN 2373-8731, Vol. 1, no 5, p. 1-6Article in journal (Refereed) Published
Abstract [en]

Background: Effective digestive enzymes are crucial for successful islet isolation. Supplemental proteases are essential as they synergize with collagenase for effective pancreas digestion. The presence of tryptic-like activity has been implicated in efficient enzyme blends and the present study aimed to evaluate if addition of clostripain, an enzyme with tryptic-like activity, could improve efficacy of the islet isolation procedure.

Methods: Clostripain was added to the enzyme blend just before pancreas perfusion. Islets were isolated per standard method and numerous isolation parameters, islet quality control, and the number of isolations fulfilling standard transplantation criteria were evaluated. Two control organs per clostripain organ were chosen by blindly matching against body mass index, cold ischemia time, hemoglobin A1c, donor sex, and donor age.

Results: There were no differences in pancreas weight, dissection time, digestion time, harvest time, percent digested pancreas, or total pellet volume before islet purification between control or clostripain pancreases. Glucose-stimulated insulin release results were similar between groups. Total isolation islet equivalents, purified tissue volume and islet equivalents/g pancreas as well as fulfillment of transplantation criteria favored clostripain processed pancreases.

Conclusions: The addition of clostripain to the enzyme blend soundly improved islet yields and transplantation rates. It gently aided pancreas digestion and maintained proper islet functionality. The addition of clostripain to the enzyme blend has now been implemented into standard isolation protocols at the isolation centers in Uppsala and in Oslo.

National Category
Endocrinology and Diabetes
Identifiers
urn:nbn:se:uu:diva-254354 (URN)10.1097/TXD.0000000000000528 (DOI)
Available from: 2015-07-30 Created: 2015-06-08 Last updated: 2015-08-28Bibliographically approved
Goto, M., Friberg, A., Ståhle, M., Imura, T., Yamagata, Y., Watanabe, K., . . . Korsgren, O. (2015). Proof of concept for the clinical application of animal component free recombinant collagenase for isolating pancreatic islets. Paper presented at IPITA/IXA/CTS Joint Congress, NOV 15-19, 2015, Melbourne, AUSTRALIA. Xenotransplantation, 22, S50-S50
Open this publication in new window or tab >>Proof of concept for the clinical application of animal component free recombinant collagenase for isolating pancreatic islets
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2015 (English)In: Xenotransplantation, ISSN 0908-665X, E-ISSN 1399-3089, Vol. 22, p. S50-S50Article in journal, Meeting abstract (Other academic) Published
National Category
Other Clinical Medicine
Identifiers
urn:nbn:se:uu:diva-276044 (URN)000364594100117 ()
Conference
IPITA/IXA/CTS Joint Congress, NOV 15-19, 2015, Melbourne, AUSTRALIA
Available from: 2016-02-09 Created: 2016-02-09 Last updated: 2017-11-30Bibliographically approved
Goto, M., Friberg, A., Ståhle, M., Imura, T., Yamagata, Y., Watanabe, K., . . . Korsgren, O. (2015). Proof Of Concept For The Clinical Application Of Animal Component Free Recombinant Collagenase For Isolating Pancreatic Islets. Paper presented at Joint Congress of the International-Pancreas-and-Islet-Transplantation-Association, International-Xenotransplantation-Association and Cell-Transplant-Society, NOV 15-19, 2015, Melbourne, AUSTRALIA. Transplantation, 99(11), S80-S80
Open this publication in new window or tab >>Proof Of Concept For The Clinical Application Of Animal Component Free Recombinant Collagenase For Isolating Pancreatic Islets
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2015 (English)In: Transplantation, ISSN 0041-1337, E-ISSN 1534-6080, Vol. 99, no 11, p. S80-S80Article in journal, Meeting abstract (Other academic) Published
National Category
Immunology in the medical area
Identifiers
urn:nbn:se:uu:diva-281551 (URN)000368625700114 ()
Conference
Joint Congress of the International-Pancreas-and-Islet-Transplantation-Association, International-Xenotransplantation-Association and Cell-Transplant-Society, NOV 15-19, 2015, Melbourne, AUSTRALIA
Available from: 2016-04-05 Created: 2016-03-24 Last updated: 2018-01-10Bibliographically approved
Ståhle, M., Friberg, A. & Korsgren, O. (2013). Commercial Enzymes for Islet Isolation: Integrity and Degradation Over Time and Temperature. Paper presented at 14th World Congress of the International-Pancreas-and-Islet-Transplant-Association (IPITA), SEP 24-27, 2013, Monterey, CA. Transplantation, 96(6), S143-S143
Open this publication in new window or tab >>Commercial Enzymes for Islet Isolation: Integrity and Degradation Over Time and Temperature
2013 (English)In: Transplantation, ISSN 0041-1337, E-ISSN 1534-6080, Vol. 96, no 6, p. S143-S143Article in journal, Meeting abstract (Other academic) Published
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-220758 (URN)000330443500257 ()
Conference
14th World Congress of the International-Pancreas-and-Islet-Transplant-Association (IPITA), SEP 24-27, 2013, Monterey, CA
Available from: 2014-03-20 Created: 2014-03-20 Last updated: 2017-12-05Bibliographically approved
Ståhle, M., Honkanen-Scott, M., Ingvast, S., Korsgren, O. & Friberg, A. S. (2013). Human islet isolation processing times shortened by one hour: minimized incubation time between tissue harvest and islet purification [Letter to the editor]. Transplantation, 96(12), e91-e93
Open this publication in new window or tab >>Human islet isolation processing times shortened by one hour: minimized incubation time between tissue harvest and islet purification
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2013 (English)In: Transplantation, ISSN 0041-1337, E-ISSN 1534-6080, Vol. 96, no 12, p. e91-e93Article in journal, Letter (Refereed) Published
National Category
Biomedical Laboratory Science/Technology
Identifiers
urn:nbn:se:uu:diva-274641 (URN)10.1097/01.TP.0000437562.31212.d5 (DOI)24342944 (PubMedID)
Available from: 2016-01-25 Created: 2016-01-25 Last updated: 2017-11-30Bibliographically approved
Ståhle, M., Honkanen-Scott, M., Ingvast, S., Korsgren, O. & Friberg, A. S. (2013). Human Islet Isolation Processing Times Shortened By One Hour: Minimized Incubation Time Between Tissue Harvest and Islet Purification [Letter to the editor]. Transplantation, 96(12), E91-E93
Open this publication in new window or tab >>Human Islet Isolation Processing Times Shortened By One Hour: Minimized Incubation Time Between Tissue Harvest and Islet Purification
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2013 (English)In: Transplantation, ISSN 0041-1337, E-ISSN 1534-6080, Vol. 96, no 12, p. E91-E93Article in journal, Letter (Refereed) Published
National Category
Endocrinology and Diabetes
Identifiers
urn:nbn:se:uu:diva-220216 (URN)10.1097/01.tp.0000437562.31212.d5 (DOI)000330439900004 ()24342944 (PubMedID)
Available from: 2014-03-17 Created: 2014-03-12 Last updated: 2017-12-05Bibliographically approved
Friberg, A. S., Ståhle, M., Honkanen-Scott, M., Ingvast, S. & Korsgren, O. (2013). Human Islet Isolation Processing Times Shortened by One Hour: No Incubation Between Tissue Harvest and Isopycnic Islet Purification. Paper presented at 14th World Congress of the International-Pancreas-and-Islet-Transplant-Association (IPITA), SEP 24-27, 2013, Monterey, CA. Transplantation, 96(6), S146-S146
Open this publication in new window or tab >>Human Islet Isolation Processing Times Shortened by One Hour: No Incubation Between Tissue Harvest and Isopycnic Islet Purification
Show others...
2013 (English)In: Transplantation, ISSN 0041-1337, E-ISSN 1534-6080, Vol. 96, no 6, p. S146-S146Article in journal, Meeting abstract (Other academic) Published
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-220749 (URN)000330443500263 ()
Conference
14th World Congress of the International-Pancreas-and-Islet-Transplant-Association (IPITA), SEP 24-27, 2013, Monterey, CA
Available from: 2014-03-24 Created: 2014-03-20 Last updated: 2017-12-05Bibliographically approved
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