title: Enzymes for Pancreatic Islet Isolation Impact Chemokine-Production and Polarization of Insulin-Producing β-Cells with Reduced Functional Survival of Immunoisolated Rat Islet-Allografts as a Consequence.
abstract: The primary aim of this study was to determine whether normal variations in enzyme-activities of collagenases applied for rat-islet isolation impact longevity of encapsulated islet grafts. Also we studied the functional and immunological properties of rat islets isolated with different enzyme preparations to determine whether this impacts these parameters. Rat-islets were isolated from the pancreas with two different collagenases with commonly accepted collagenase, neutral protease, and clostripain activities. Islets had a similar and acceptable glucose-induced insulin-release profile but a profound statistical significant difference in production of the chemokines IP-10 and Gro-α. The islets were studied with nanotomy which is an EM-based technology for unbiased study of ultrastructural features of islets such as cell-cell contacts, endocrine-cell condition, ER stress, mitochondrial conditions, and cell polarization. The islet-batch with higher chemokine-production had a lower amount of polarized insulin-producing β-cells. All islets had more intercellular spaces and less interconnected areas with tight cell-cell junctions when compared to islets in the pancreas. Islet-graft function was studied by implanting encapsulated and free islet grafts in rat recipients. Alginate-based encapsulated grafts isolated with the enzyme-lot inducing higher chemokine production and lower polarization survived for a two-fold shorter period of time. The lower survival-time of the encapsulated grafts was correlated with a higher influx of inflammatory cells at 7 days after implantation. Islets from the same two batches transplanted as free unencapsulated-graft, did not show any difference in survival or function in vivo. Lack of insight in factors contributing to the current lab-to-lab variation in longevity of encapsulated islet-grafts is considered to be a threat for clinical application. Our data suggest that seemingly minor variations in activity of enzymes applied for islet-isolation might contribute to longevity-variations of immunoisolated islet-grafts.
authors.affiliation: Immunoendocrinology, department of Pathology and Medical biology, University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
authors.forename: Paul
authors.initials: P
authors.lastname: de Vos
authors.affiliation: Immunoendocrinology, department of Pathology and Medical biology, University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
authors.forename: Alexandra M
authors.initials: AM
authors.lastname: Smink
authors.affiliation: Immunoendocrinology, department of Pathology and Medical biology, University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
authors.forename: Genaro
authors.initials: G
authors.lastname: Paredes
authors.affiliation: Department of Surgery and Biomedical Engineering, University of California Irvine, Orange, CA, 92868, United States of America.
authors.forename: Jonathan R T
authors.initials: JR
authors.lastname: Lakey
authors.affiliation: Department of Cell Biology, University Medical Center Groningen, University of Groningen, P. O. Box 196, 9700 AD, Groningen, The Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Kuipers
authors.affiliation: Department of Cell Biology, University Medical Center Groningen, University of Groningen, P. O. Box 196, 9700 AD, Groningen, The Netherlands.
authors.forename: Ben N G
authors.initials: BN
authors.lastname: Giepmans
authors.affiliation: Immunoendocrinology, department of Pathology and Medical biology, University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
authors.forename: Bart J
authors.initials: BJ
authors.lastname: de Haan
authors.affiliation: Immunoendocrinology, department of Pathology and Medical biology, University of Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands.
authors.forename: Marijke M
authors.initials: MM
authors.lastname: Faas
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D016764:Cell Polarity; D002469:Cell Separation / Q000379:methods*; D018914:Cells, Immobilized; D054360:Chemokine CXCL1 / Q000096:biosynthesis / Q000276:immunology; D054357:Chemokine CXCL10 / Q000096:biosynthesis / Q000276:immunology; D017364:Collagenases / Q000737:chemistry; D003546:Cysteine Endopeptidases / Q000737:chemistry; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D004721:Endoplasmic Reticulum / Q000276:immunology / Q000648:ultrastructure; D006085:Graft Survival; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000276:immunology; D007365:Intercellular Junctions / Q000276:immunology / Q000648:ultrastructure; D007515:Islets of Langerhans / Q000166:cytology* / Q000276:immunology; D016381:Islets of Langerhans Transplantation; D008297:Male; D008928:Mitochondria / Q000276:immunology / Q000648:ultrastructure; D010447:Peptide Hydrolases / Q000737:chemistry; D051381:Rats; D013311:Streptozocin; D014184:Transplantation, Homologous
chemical_list: D001786:Blood Glucose; D054360:Chemokine CXCL1; D054357:Chemokine CXCL10; C091326:Cxcl10 protein, rat; D013311:Streptozocin; D010447:Peptide Hydrolases; D003546:Cysteine Endopeptidases; C019913:clostripain; D017364:Collagenases
references.citation: Diabetes. 1999 Jul;48(7):1381-8
references.pmid: 10389842
references.citation: Diabetologia. 2001 May;44(5):646-53
references.pmid: 11380084
references.citation: J Immunol. 2002 Apr 1;168(7):3195-204
references.pmid: 11907072
references.citation: Biotechniques. 2002 Mar;32(3):612-4, 616, 618-9
references.pmid: 11911663
references.citation: J Biomed Mater Res. 2002 Dec 5;62(3):430-7
references.pmid: 12209929
references.citation: Biomaterials. 2003 Jan;24(2):305-12
references.pmid: 12419632
references.citation: Xenotransplantation. 2003 May;10(3):240-51
references.pmid: 12694544
references.citation: Diabetologia. 2003 May;46(5):666-73
references.pmid: 12750768
references.citation: Transpl Int. 2003 Sep;16(9):613-32
references.pmid: 12928769
references.citation: J Surg Res. 1992 Nov;53(5):502-9
references.pmid: 1434601
references.citation: Diabetologia. 1992 Oct;35(10):917-23
references.pmid: 1451947
references.citation: Cell Transplant. 2003;12(6):617-25
references.pmid: 14579930
references.citation: J Biomed Mater Res A. 2003 Dec 15;67(4):1219-27
references.pmid: 14624508
references.citation: Biotechnol Appl Biochem. 2004 Apr;39(Pt 2):159-64
references.pmid: 15032736
references.citation: Pancreas. 2004 Jul;29(1):e15-22
references.pmid: 15211119
references.citation: Cell Transplant. 2004;13(5):515-24
references.pmid: 15565864
references.citation: Transpl Immunol. 2004 Dec;13(4):289-96
references.pmid: 15589742
references.citation: Diabetologia. 2005 May;48(5):906-12
references.pmid: 15830183
references.citation: J Biomed Mater Res A. 2006 Feb;76(2):243-51
references.pmid: 16265647
references.citation: Transplantation. 2006 May 15;81(9):1345-53
references.pmid: 16699465
references.citation: Biomaterials. 2006 Oct;27(28):4831-9
references.pmid: 16766026
references.citation: J Biomed Mater Res A. 2007 Mar 15;80(4):813-9
references.pmid: 17058213
references.citation: Transplantation. 2007 Jan 15;83(1):7-12
references.pmid: 17220782
references.citation: Am J Transplant. 2007 May;7(5):1233-41
references.pmid: 17359501
references.citation: Diabetologia. 2008 Feb;51(2):227-32
references.pmid: 18040664
references.citation: Transplantation. 1991 Nov;52(5):777-83
references.pmid: 1835193
references.citation: J Biomed Mater Res A. 2009 Jun;89(3):609-15
references.pmid: 18435412
references.citation: Arterioscler Thromb Vasc Biol. 2008 Nov;28(11):1950-9
references.pmid: 18818421
references.citation: Biomaterials. 2009 May;30(13):2559-70
references.pmid: 19201460
references.citation: Curr Opin Organ Transplant. 2010 Feb;15(1):79-85
references.pmid: 19907327
references.citation: Adv Exp Med Biol. 2010;654:627-40
references.pmid: 20217517
references.citation: Adv Exp Med Biol. 2010;670:5-21
references.pmid: 20384214
references.citation: Cell Transplant. 2010;19(8):1031-46
references.pmid: 20546673
references.citation: Mol Cell Biol. 1990 Oct;10(10):5596-9
references.pmid: 2078213
references.citation: Transplantation. 2011 Jan 27;91(2):137-45
references.pmid: 21116222
references.citation: Acta Biomater. 2011 Apr;7(4):1683-92
references.pmid: 21145438
references.citation: Curr Diab Rep. 2011 Oct;11(5):384-91
references.pmid: 21826429
references.citation: J Microencapsul. 2011;28(6):499-507
references.pmid: 21827357
references.citation: Curr Diab Rep. 2011 Oct;11(5):345-54
references.pmid: 21830042
references.citation: Diabetologia. 2011 Oct;54(10):2494-505
references.pmid: 21830149
references.citation: Proc Natl Acad Sci U S A. 2012 Mar 27;109(13):5022-7
references.pmid: 22393012
references.citation: Cell Transplant. 2012;21(9):2063-78
references.pmid: 22404979
references.citation: Expert Opin Biol Ther. 2012 Jun;12(6):731-41
references.pmid: 22506840
references.citation: Biomaterials. 2012 Aug;33(22):5552-9
references.pmid: 22560199
references.citation: Cell Transplant. 2012;21(2-3):465-71
references.pmid: 22793054
references.citation: Cell Transplant. 2012;21(2-3):501-8
references.pmid: 22793058
references.citation: Cell Transplant. 2013;22(8):1463-76
references.pmid: 23043896
references.citation: PLoS One. 2013;8(2):e56924
references.pmid: 23437272
references.citation: Sci Rep. 2013;3:1804
references.pmid: 23652855
references.citation: Adv Drug Deliv Rev. 2014 Apr;67-68:111-30
references.pmid: 23876549
references.citation: J Control Release. 2013 Dec 28;172(3):983-92
references.pmid: 24051034
references.citation: Adv Drug Deliv Rev. 2014 Apr;67-68:15-34
references.pmid: 24270009
references.citation: Nat Med. 2014 Mar;20(3):233
references.pmid: 24603789
references.citation: Front Bioeng Biotechnol. 2014 Aug 06;2:26
references.pmid: 25147785
references.citation: PLoS One. 2014 Oct 27;9(10):e109837
references.pmid: 25347191
references.citation: Islets. 2014;6(4):e977118
references.pmid: 25437379
references.citation: Trends Pharmacol Sci. 2015 Aug;36(8):537-46
references.pmid: 26067102
references.citation: Materials (Basel). 2014 Mar 12;7(3):2087-2103
references.pmid: 28788557
references.citation: J Exp Med. 1995 Jul 1;182(1):155-62
references.pmid: 7540647
references.citation: Transpl Int. 1994 Jul;7(4):264-71
references.pmid: 7916926
references.citation: Transplantation. 1994 Apr 15;57(7):1010-7
references.pmid: 8165696
references.citation: Transplant Proc. 1994 Apr;26(2):641-2
references.pmid: 8171590
references.citation: Cell Transplant. 1996 Jan-Feb;5(1):21-30
references.pmid: 8665073
references.citation: J Clin Invest. 1996 Sep 15;98(6):1417-22
references.pmid: 8823307
references.citation: Transplantation. 1996 Oct 15;62(7):888-93
references.pmid: 8878379
references.citation: Transplantation. 1996 Oct 15;62(7):893-9
references.pmid: 8878380
references.citation: Diabetologia. 1997 Mar;40(3):262-70
references.pmid: 9084963
references.citation: Transplantation. 1997 Mar 27;63(6):824-30
references.pmid: 9089221
references.citation: Transplant Proc. 1997 Feb-Mar;29(1-2):756-7
references.pmid: 9123511
references.citation: Biomaterials. 1997 Aug;18(16):1085-90
references.pmid: 9247345