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Horm Metab Res 2003; 35(8): 460-465
DOI: 10.1055/s-2003-41802
Original Basic
© Georg Thieme Verlag Stuttgart · New York

Beneficial Effect Of Pretreatment Of Islets With Fibronectin On Glucose Tolerance After Islet Transplantation

Y.  Hamamoto1, 2 , S.  Fujimoto1 , A.  Inada1, 2 , M.  Takehiro1 , K.  Nabe1 , D.  Shimono1 , M.  Kajikawa1 , J.  Fujita1 , Y.  Yamada1 , Y.  Seino1
  • 1Department of Metabolism and Clinical Nutrition, Graduate School of Medicine, Kyoto University, Japan
  • 2Joslin Diabetes Center, Harvard Medical School, Section of Islet Transplantation and Cell Biology, Boston, MA, USA
Further Information

Publication History

Received 9 December 2002

Accepted after revision 7 April 2003

Publication Date:
02 September 2003 (online)

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Abstract

The scarcity of available islets is an obstacle for clinically successful islet transplantation. One solution might be to increase the efficacy of the limited islets. Isolated islets are exposed to a variety of cellular stressors, and disruption of the cell-matrix connections damages islets. We examined the effect of fibronectin, a major component of the extracellular matrix, on islet viability, mass and function, and also examined whether fibronectin-treated islets improved the results of islet transplantation. Islets cultured with fibronectin for 48 hours maintained higher cell viability (0.146 ± 0.010 vs. 0.173 ± 0.007 by MTT assay), and also had a greater insulin and DNA content (86.8 ± 3.6 vs. 72.8 ± 3.2 ng/islet and 35.2 ± 1.4 vs. 30.0 ± 1.5 ng/islet, respectively) than islets cultured without fibronectin (control). Absolute values of insulin secretion were higher in fibronectin-treated islets than in controls; however, the ratio of stimulated insulin secretion to basal secretion was not significantly different (206.9 ± 23.3 vs. 191.7 ± 20.2 % when the insulin response to 16.7 mmol/l glucose was compared to that of 3.3 mmol/l glucose); the higher insulin secretion was thus mainly due to larger islet cell mass. The rats transplanted with fibronectin-treated islets had lower plasma glucose and higher plasma insulin levels within 2 weeks after transplantation, and had more favorable glucose tolerance 9 weeks after transplantation. These results indicate that cultivation with fibronectin might preserve islet cell viability, mass and insulin secretory function, which could improve glucose tolerance following islet transplantation.

Key words

Extracellular matrix - Cell mass - DNA content - Insulin content - Insulin secretion

References

  • 1 Shapiro A MJ, Lakey J RT, Ryan E A, Korbutt G S, Toth E, Warnock G L, Kneteman N M, Rajotte R V. Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid - free immunosuppressive regimen.  N Engl J Med. 2000;  343 230-238
    CrossrefPubMedSearch in Google Scholar
  • 2 Rosenberg L, Wang R, Paraskevas S, Maysinger D. Structural and functional changes resulting from islet isolation lead to islet cell death.  Surgery. 1999;  126 393-398
    CrossrefPubMedSearch in Google Scholar
  • 3 Meredith J E, Fazeli B, Schwartz M A. The extracellular matrix as a cell survival factor.  Mol Biol Cell. 1993;  4 953-961
    PubMedSearch in Google Scholar
  • 4 Brendel M D, Kong S S, Alejandro R, Mintz D H. Improved functional survival of human islets of Langerhans in three - dimensional matrix culture.  Cell Transplant. 1994;  3 427-435
    PubMedSearch in Google Scholar
  • 5 Jiang F X, Cram D S, DeAizpurua H J, Harrison L C. Laminin-1 promotes differentiation of fetal mouse pancreatic beta-cells.  Diabetes. 1999;  48 722-730
    PubMedSearch in Google Scholar
  • 6 Ilieva A, Yuan S, Wang R N, Agapitos D, Hill D J, Rosenberg L. Pancreatic islet cell survival following islet isolation: the role of cellular interactions in the pancreas.  J Endocrinol. 1999;  161 357-364
    CrossrefPubMedSearch in Google Scholar
  • 7 Wang R N, Rosenberg L. Maintenance of beta-cell function and survival following islet isolation requires re-establishment of the islet-matrix relationship.  J Endocrinol. 1999;  163 181-190
    CrossrefPubMedSearch in Google Scholar
  • 8 Nagata N, Gu Y, Hori H, Balamurugan A N, Touma M, Kawakami Y, Wang W, Baba T T, Satake A, Nozawa M, Tabata Y, Inoue K. Evaluation of insulin secretion of isolated rat islets cultured in extracellular matrix.  Cell Transplant. 2001;  10 447-451
    PubMedSearch in Google Scholar
  • 9 Frisch S M, Francis H. Disruption of epithelial cell-matrix interactions induces apoptosis.  J Cell Biol. 1994;  124 619-626
    CrossrefPubMedSearch in Google Scholar
  • 10 Thomas F T, Contreras J L, Bilbao G, Ricordi C, Curiel D, Thomas J M. Anoikis, extracellular matrix, and apoptosis factors in isolated cell transplantation.  Surgery. 1999;  126 299-304
    CrossrefPubMedSearch in Google Scholar
  • 11 Paraskevas S, Maysinger D, Wang R, Duguid W P, Rosenberg L. Cell loss in isolated human islets occurs by apoptosis.  Pancreas. 2000;  20 270-276
    CrossrefPubMedSearch in Google Scholar
  • 12 Cattan P, Berney T, Schena S, Molano D, Pileggi A, Vizzardelli C, Ricordi C, Inverardi L. Early assessment of apoptosis in isolated islets of Langerhans.  Transplantation. 2001;  71 857-862
    PubMedSearch in Google Scholar
  • 13 Frisch S M, Rouslahti E. Integrins and anoikis.  Curr Opin Cell Biol. 1997;  9 701-706
    CrossrefPubMedSearch in Google Scholar
  • 14 Thomas F, Wu J, Contreras J L, Smyth C, Bilbao G, He J, Thomas J. A tripartite anoikis-like mechanism causes early isolated islet apoptosis.  Surgery. 2001;  130 333-338
    CrossrefPubMedSearch in Google Scholar
  • 15 Zhang Z, Vuori K, Reed J C, Rouslahti E. The α5β1 integrin supports survival of cells on fibronectin and up-regulates Bcl-2 expression.  Proc Natl Acad Sci. 1995;  92 6161-6165
    PubMedSearch in Google Scholar
  • 16 de la Fuente M T, Casanova B, Garcia-Gila M, Silva A, Garcia-Pardo A. Fibronectin interaction with α4β1 integrin prevents apoptosis in B cell chronic lymphocytic leukemia: correlation with Bcl-2 and Bax.  Leukemia. 1999;  13 266-274
    CrossrefPubMedSearch in Google Scholar
  • 17 Anwar A RF, Moqbel R, Walsh G M, Kay A B, Wardlaw A J. Adhesion to fibronectin prolongs eosinophil survival.  J Exp Med. 1993;  177 839-843
    CrossrefPubMedSearch in Google Scholar
  • 18 Wang R N, Paraskevas S, Rosenberg L. Characterization of integrin in islets isolated from Hamster, Canine, Porcine, and Human pancreas.  J Histochem Cytochem. 1999;  47 499-506
    PubMedSearch in Google Scholar
  • 19 Bosco D, Meda P, Halben P A, Rouiller D G. Importance of cell - matrix interactions in rat islet β-cell secretion in vitro. Role of α6β1 integrin.  Diabetes. 2000;  49 233-243
    CrossrefPubMedSearch in Google Scholar
  • 20 Sutton R, Peters M, Mcshane P, Gray D WR, Morris P J. Isolation of rat pancreatic islets by ductal injection of collagenase.  Transplantation. 1986;  42 689-691
    CrossrefPubMedSearch in Google Scholar
  • 21 Tsuura Y, Ishida H, Okamoto Y, Kato S, Horie M, Ikeda H, Seino Y. Reduced sensitivity of dihydroxyacetone on ATP-sensitive K+ channels of pancreatic beta-cells in GK rats.  Diabetologia. 1994;  37 1082-1087
    PubMedSearch in Google Scholar
  • 22 Fujimoto S, Tsuura Y, Ishida H, Tsuji K, Mukai E, Kajikawa M, Hamamoto Y, Takeda T, Yamada Y, Seino Y. Augmentation of basal insulin release from rat islets by preexposure to a high concentration of glucose.  Am J Physiol. 2000;  279 E927-940
    PubMedSearch in Google Scholar
  • 23 Hopcroft D W, Mason D R, Scott R S. Standardization of insulin secretion from pancreatic islets: Validation of a DNA assay.  Horm Metab Res. 1985;  17 559-561
    PubMedSearch in Google Scholar
  • 24 Kersgren O, Jansson L, Andersson A. Effects of hyperglycemia on function of isolated mouse pancreatic islets transplanted under kidney capsule.  Diabetes. 1989;  38 510-515
    PubMedSearch in Google Scholar
  • 25 Hamamoto Y, Tsuura Y, Fujimoto S, Nagata M, Takeda T, Mukai E, Fujita J, Yamada Y, Seino Y. Recovery of function and mass of endogenous β-cells in streptozotocin - induced diabetic rats treated with islet transplantation.  Biochem Biophys Res Commun. 2001;  287 104-109
    CrossrefPubMedSearch in Google Scholar
  • 26 Ricordi C. Pancreatic islet cell transplantation; 1892 - 1992 One century of transplantation for diabetes. Austin; R. G. Landes Co 1992
    Search in Google Scholar
  • 27 Shapiro A M, Hao E, Rajotte R V, Kneteman N M. High yield of rodent islets with intraductal collagenase and stationary digestion - a comparison with standard technique.  Cell Transplant. 1996;  6 631-638
    PubMedSearch in Google Scholar
  • 28 Ranuncoli A, Cautero N, Ricordi C, Masetti M, Malono R D, Inverardi L, Alejandro R, Kenyon N S. Islet cell transplantation: in vivo and in vitro functional assessment of nonhuman primate pancreatic islets.  Cell Transplant. 2000;  9 409-414
    PubMedSearch in Google Scholar
  • 29 Ricordi C, Lakey J RT, Hering B J. Challenges toward standardization of islet isolation technology.  Transplant Proc. 2001;  33 1709
    CrossrefPubMedSearch in Google Scholar
  • 30 Pileggi A, Ricordi C, Alessiani M, Inverardi L. Factors influencing islet of Langerhans graft function and monitoring.  Clin Chim Acta. 2001;  310 3-16
    CrossrefPubMedSearch in Google Scholar
  • 31 Munn S R, Kaufman D B, Field M J, Sutherland D E. Successful 24- and 48-hour cold storage of the pancreas prior to islet isolation and transplantation.  Transplant Proc. 1989;  21 2635-2637
    PubMedSearch in Google Scholar
  • 32 Hesse U J, Berrevoet P, Pattyn P, Vanholder R, de Hemptinne B. Donor parameters of pancreas grafts processed for islet or β-cell isolation and transplantation.  Transplant Proc. 1997;  29 2259
    CrossrefPubMedSearch in Google Scholar
  • 33 Perfetti R, Henderson T E, Wang Y, Montrose-Rafizadeh C, Egan J M. Insulin release and insulin mRNA levels in rat islets of Langerhans cultured on extracellular matrix.  Pancreas. 1996;  13 47-54
    PubMedSearch in Google Scholar
  • 34 Bosco D, Gonelle-Gispert C, Wollheim C B, Halben P A, Rouiller D G. Increased intracellular calcium is required for spreading of rat islet β-cells on extracellular matrix.  Diabetes. 2001;  50 1039-1046
    PubMedSearch in Google Scholar
  • 35 Davalli A M, Scaglia L, Zangen D H, Hollister J, Bonnar-Weir S, Weir G C. Vulnerability of islets in the immediate posttransplantation period.  Diabetes. 1996;  19 1161-1167
    PubMedSearch in Google Scholar
  • 36 Menger M D, Yamauchi J, Vollmar B. Revascularization and microcirculation of freely grafted islets of Langerhans.  World J Surg. 2001;  25 509-515
    CrossrefPubMedSearch in Google Scholar
  • 37 Ryan E A, Lakey J RT, Paty B W, Imes S, Korbutt G S, Kneteman N M, Bigam D, Rajotte R V, Shapiro A MJ. Successful islet transplantation. Continued insulin reserve provides long-term glycemic control.  Diabetes. 2002;  51 2148-2157
    CrossrefPubMedSearch in Google Scholar
  • 38 Laybutt D R, Glandt M, Xu G, Ahn Y B, Trivedi N, Bonner-Weir S, Weir G C. Critical reduction in β-cell mass results in two distinct outcomes over time.  J Biol Chem. 2003;  278 2997-3005
    CrossrefPubMedSearch in Google Scholar

Y. Hamamoto, M.D. PhD. 

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