Pancreas collagen digestion during islet of Langerhans isolation—a prospective study

The success of pancreas islet isolation largely depends on donor characteristics, including extracellular matrix composition of which collagen is the main element. We hypothesized that isolation yields are proportional to collagen digestion percentage, and aimed to determine a threshold that predicts isolation success. The amount of pancreas collagen (I‐V) was determined using colorimetry prior to and after the digestion process in 52 human islet isolations. Collagen I‐V and VI were also assessed histologically. We identified a collagen digestion threshold of ≥ 60% as an independent factor beyond which an islet preparation has a ninefold increased odds of yielding ≥ 250 000 islet equivalents (IEQ) (P = 0.009) and a sixfold increased odds of being transplanted (P = 0.015). Preparations with ≥ 60% collagen digestion (n = 35) yielded 283 017 ± 164 214 IEQ versus 180 142 ± 85 397 in the < 60% collagen digestion group (n = 17) (P = 0.016); respectively 62.9% versus 29.4% of those were transplanted (P = 0.024). Common donor characteristics, initial collagen content, enzyme blend, and digestion times were not associated with collagen digestion percentage variations. Donor age positively correlated with the amount of collagen VI (P = 0.013). There was no difference in islet graft survival between high and low digestion groups. We determined that a 60% pancreas collagen digestion is the threshold beyond which an islet isolation is likely to be successful and transplanted.

[1]  Lingjun Li,et al.  In-depth quantification of extracellular matrix proteins from human pancreas. , 2019, Journal of proteome research.

[2]  T. Berney,et al.  Pancreas preservation fluid microbial contamination is associated with poor islet isolation outcomes – a multi‐centre cohort study , 2018, Transplant international : official journal of the European Society for Organ Transplantation.

[3]  Xia Li,et al.  Extracellular matrix scaffold and hydrogel derived from decellularized and delipidized human pancreas , 2018, Scientific Reports.

[4]  C. Wandrey,et al.  Multipotent mesenchymal stromal cells enhance insulin secretion from human islets via N-cadherin interaction and prolong function of transplanted encapsulated islets in mice , 2017, Stem cell research & therapeutics.

[5]  S. Cross,et al.  &bgr; Cell Replacement Therapy: The Next 10 Years , 2017, Transplantation.

[6]  P. Rorsman,et al.  Key Matrix Proteins Within the Pancreatic Islet Basement Membrane Are Differentially Digested During Human Islet Isolation , 2017, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[7]  S. Strom,et al.  Current status of hepatocyte xenotransplantation. , 2015, International journal of surgery.

[8]  T. Berney,et al.  Five-Year Metabolic, Functional, and Safety Results of Patients With Type 1 Diabetes Transplanted With Allogenic Islets Within the Swiss-French GRAGIL Network , 2015, Diabetes Care.

[9]  M. Rickels,et al.  Islet Product Characteristics and Factors Related to Successful Human Islet Transplantation From the Collaborative Islet Transplant Registry (CITR) 1999–2010 , 2014, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[10]  T. Berney,et al.  Islet of Langerhans isolation from pediatric and juvenile donor pancreases , 2014, Transplant international : official journal of the European Society for Organ Transplantation.

[11]  T. Berney,et al.  Survival of Free and Encapsulated Human and Rat Islet Xenografts Transplanted into the Mouse Bone Marrow , 2014, PloS one.

[12]  P. Morel,et al.  Transplantation of mesenchymal stem cells for the treatment of liver diseases, is there enough evidence? , 2013, Stem cell research.

[13]  Nicole A. Turgeon,et al.  Improvement in Outcomes of Clinical Islet Transplantation: 1999–2010 , 2012, Diabetes Care.

[14]  F. Ris,et al.  Comparative Impact on Islet Isolation and Transplant Outcome of the Preservation Solutions Institut Georges Lopez-1, University of Wisconsin, and Celsior , 2012, Transplantation.

[15]  T. Berney,et al.  Impact of the Number of Infusions on 2-Year Results of Islet-After-Kidney Transplantation in the GRAGIL Network , 2011, Transplantation.

[16]  T. Berney,et al.  Cadherin engagement protects human β-cells from apoptosis. , 2011, Endocrinology.

[17]  F. Ris,et al.  Islet Autotransplantation After Extended Pancreatectomy for Focal Benign Disease of the Pancreas , 2011, Transplantation.

[18]  T. Berney,et al.  Influence of Donor Age on Islet Isolation and Transplantation Outcome , 2010, Transplantation.

[19]  O. Terpstra,et al.  Porcine islet isolation outcome is not affected by the amount and distribution of collagen in the pancreas , 2010, Xenotransplantation.

[20]  M. Tabrizian,et al.  The effect of extracellular matrix components on the preservation of human islet function in vitro. , 2010, Biomaterials.

[21]  S. Totey,et al.  Open-labeled study of unilateral autologous bone-marrow-derived mesenchymal stem cell transplantation in Parkinson's disease. , 2010, Translational research : the journal of laboratory and clinical medicine.

[22]  G. Szot,et al.  Successful clinical islet isolation using a GMP-manufactured collagenase and neutral protease. , 2009, Transplantation.

[23]  E. Renard,et al.  Quality of life after islet transplantation: data from the GRAGIL 1 and 2 trials , 2009, Diabetic medicine : a journal of the British Diabetic Association.

[24]  J. Stendahl,et al.  Extracellular Matrix in Pancreatic Islets: Relevance to Scaffold Design and Transplantation , 2009, Cell transplantation.

[25]  Philippe Morel,et al.  Computer-Assisted Digital Image Analysis to Quantify the Mass and Purity of Isolated Human Islets Before Transplantation , 2008, Transplantation.

[26]  E. Renard,et al.  Expectations and Strategies Regarding Islet Transplantation: Metabolic Data From the GRAGIL 2 Trial , 2007, Transplantation.

[27]  Philippe Morel,et al.  International trial of the Edmonton protocol for islet transplantation. , 2006, The New England journal of medicine.

[28]  T. Berney,et al.  Sequential Kidney/Islet Transplantation: Efficacy and Safety Assessment of a Steroid‐Free Immunosuppression Protocol , 2006, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[29]  S. Hughes,et al.  Characterisation of Collagen VI within the Islet-Exocrine Interface of the Human Pancreas: Implications for Clinical Islet Isolation? , 2006, Transplantation.

[30]  Rennian Wang,et al.  Role for β1 Integrin and Its Associated α3, α5, and α6 Subunits in Development of the Human Fetal Pancreas , 2005 .

[31]  T. Berney,et al.  Assessment of a Novel Two-Component Enzyme Preparation for Human Islet Isolation and Transplantation , 2005, Transplantation.

[32]  T. Berney,et al.  LOGISTICS AND TRANSPLANT COORDINATION ACTIVITY IN THE GRAGIL SWISS-FRENCH MULTICENTER NETWORK OF ISLET TRANSPLANTATION , 2004, Transplantation.

[33]  M. Rubart,et al.  Myocyte and myogenic stem cell transplantation in the heart. , 2003, Cardiovascular research.

[34]  F. Ris,et al.  Human islet transplantation network for the treatment of Type I diabetes: first data from the Swiss-French GRAGIL consortium (1999–2000) , 2001, Diabetologia.

[35]  N. Hakim.  Recent developments and future prospects in pancreatic and islet transplantation , 2001, Diabetes, obesity & metabolism.

[36]  E. Ryan,et al.  Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. , 2000, The New England journal of medicine.

[37]  S. Paraskevas,et al.  Characterization of Integrin Expression in Islets Isolated from Hamster, Canine, Porcine, and Human Pancreas , 1999, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[38]  R. van Schilfgaarde,et al.  Distribution of collagens type I, type III and type V in the pancreas of rat, dog, pig and man , 1994, Cell and Tissue Research.

[39]  Peter J. Morris,et al.  Islet isolation assessment in man and large animals , 1990, Acta diabetologia latina.

[40]  Camillo Ricordi,et al.  Automated Method for Isolation of Human Pancreatic Islets , 1988, Diabetes.

[41]  H. Seers,et al.  OUTCOME , 1977, How to Win Your Case.

[42]  T. Berney,et al.  Beta-Cell Replacement: Pancreas and Islet Cell Transplantation. , 2016, Endocrine development.

[43]  Rennian Wang,et al.  Role for beta1 integrin and its associated alpha3, alpha5, and alpha6 subunits in development of the human fetal pancreas. , 2005, Diabetes.

[44]  C. Hulstaert,et al.  Significance of the peri-insular extracellular matrix for islet isolation from the pancreas of rat, dog, pig, and man , 2004, Cell and Tissue Research.