Comprehensive Analysis of Gene Expression of Isolated Pancreatic Islets During Pretransplant Culture

Overview

Journal Fukushima J Med Sci

Specialty General Medicine

Date 2021 Feb 18

PMID 33597316

Citations 1

Authors

Junichiro Haga

Naoya Sato

Takayuki Anazawa

Takashi Kimura

Akira Kenjo

Mitsukazu Gotoh

Shigeru Marubashi

Affiliations

Soon will be listed here.

Abstract

Background: The aim of this study was to investigate the effect of pretransplant culture on the survival of pancreatic islet grafts, and to determine the biological characteristics of isolated islets during pretransplant culture.

Methods: The survival of islets from Wistar rats, transplanted to diabetic C57BL/B6 mice, was compared between fresh islets and cultured islets. A comprehensive gene expression analysis was employed to investigate biological processes during pretransplant culture, and in vitro validation studies were performed.

Results: Survival of cultured xenografts was significantly prolonged as compared to that of fresh islets (fresh:12.5 ± 1.9 days, 1-day cultured:16.0 ± 1.3 days (p= 0.017), 3-day cultured:17.0 ± 2.6 days (p= 0.014)). Comprehensive gene expression analysis identified significant upregulation of annotated functions associated with inflammation in cultured groups. Six proinflammatory genes, including heme oxygenase 1 (HO-1) and IL-6, were significantly upregulated during culture.　Validation studies revealed significantly higher levels of IL-6 in the supernatant of cultured islets and HO-1 in the cultured islets when compared with fresh islets.

Conclusion: Transplantation of cultured islets induced significant but minimal prolongation of graft survival in xenogeneic combinations. Comprehensive analysis of gene expression in cultured islets showed biological processes associated with proinflammation during culture.

Citing Articles

Induction of Immune Tolerance in Islet Transplantation Using Apoptotic Donor Leukocytes.

Sato N, Marubashi S J Clin Med. 2021; 10(22).

PMID: 34830586 PMC: 8625503. DOI: 10.3390/jcm10225306.

References

Moberg L, Johansson H, Lukinius A, Berne C, Foss A, Kallen R . Production of tissue factor by pancreatic islet cells as a trigger of detrimental thrombotic reactions in clinical islet transplantation. Lancet. 2002; 360(9350):2039-45. DOI: 10.1016/s0140-6736(02)12020-4. View

Kanak M, Takita M, Kunnathodi F, Lawrence M, Levy M, Naziruddin B . Inflammatory response in islet transplantation. Int J Endocrinol. 2014; 2014:451035. PMC: 4021753. DOI: 10.1155/2014/451035. View

Ihm S, Matsumoto I, Zhang H, Ansite J, Hering B . Effect of short-term culture on functional and stress-related parameters in isolated human islets. Transpl Int. 2008; 22(2):207-16. DOI: 10.1111/j.1432-2277.2008.00769.x. View

Gruessner R, Sutherland D, Troppmann C, Benedetti E, Hakim N, Dunn D . The surgical risk of pancreas transplantation in the cyclosporine era: an overview. J Am Coll Surg. 1997; 185(2):128-44. DOI: 10.1016/s1072-7515(01)00895-x. View

Gotoh M, Maki T, Kiyoizumi T, Satomi S, Monaco A . An improved method for isolation of mouse pancreatic islets. Transplantation. 1985; 40(4):437-8. DOI: 10.1097/00007890-198510000-00018. View

Bank H . Rapid assessment of islet viability with acridine orange and propidium iodide. In Vitro Cell Dev Biol. 1988; 24(4):266-73. DOI: 10.1007/BF02628826. View

Jahr H, Hussmann B, Eckhardt T, Bretzel R . Successful single donor islet allotransplantation in the streptozotocin diabetes rat model. Cell Transplant. 2002; 11(6):513-8. View

Ricordi C, Gray D, Hering B, Kaufman D, Warnock G, Kneteman N . Islet isolation assessment in man and large animals. Acta Diabetol Lat. 1990; 27(3):185-95. DOI: 10.1007/BF02581331. View

Itoh T, Nitta T, Nishinakamura H, Kojima D, Mera T, Ono J . HMGB1-Mediated Early Loss of Transplanted Islets Is Prevented by Anti-IL-6R Antibody in Mice. Pancreas. 2014; 44(1):166-71. DOI: 10.1097/MPA.0000000000000188. View

10.

Kedinger M, Haffen K, Grenier J, Eloy R . In vitro culture reduces immunogenicity of pancreatic endocrine islets. Nature. 1977; 270(5639):736-8. DOI: 10.1038/270736a0. View

11.

Matsuyama S, Gunji T, Ise K, Sato Y, Saito T, Gotoh M . Permanent acceptance of mitomycin C-treated islet allograft. Transplantation. 2003; 76(1):65-71. DOI: 10.1097/01.TP.0000069232.48055.03. View

12.

Kin T, Senior P, OGorman D, Richer B, Salam A, Shapiro A . Risk factors for islet loss during culture prior to transplantation. Transpl Int. 2008; 21(11):1029-35. DOI: 10.1111/j.1432-2277.2008.00719.x. View

13.

Singh A, Ramachandran S, Graham M, Daneshmandi S, Heller D, Suarez-Pinzon W . Long-term tolerance of islet allografts in nonhuman primates induced by apoptotic donor leukocytes. Nat Commun. 2019; 10(1):3495. PMC: 6677762. DOI: 10.1038/s41467-019-11338-y. View

14.

Malejczyk M, Malejczyk J, Abgarowicz K, Smogorzewski M . Immunogenicity of allogeneic pancreatic islet grafts: the effect of in vitro culture and the site of transplantation. Arch Immunol Ther Exp (Warsz). 1989; 37(1-2):11-6. View

15.

Benhamou P, Oberholzer J, Toso C, Kessler L, Penfornis A, Bayle F . Human islet transplantation network for the treatment of Type I diabetes: first data from the Swiss-French GRAGIL consortium (1999-2000). Groupe de Recherche Rhin Rhĵne Alpes Genève pour la transplantation d'Ilots de Langerhans. Diabetologia. 2001; 44(7):859-64. DOI: 10.1007/s001250100571. View

16.

Saito T, Ohashi K, Utoh R, Shimizu H, Ise K, Suzuki H . Reversal of diabetes by the creation of neo-islet tissues into a subcutaneous site using islet cell sheets. Transplantation. 2011; 92(11):1231-6. DOI: 10.1097/TP.0b013e3182375835. View

17.

Choi A, Alam J . Heme oxygenase-1: function, regulation, and implication of a novel stress-inducible protein in oxidant-induced lung injury. Am J Respir Cell Mol Biol. 1996; 15(1):9-19. DOI: 10.1165/ajrcmb.15.1.8679227. View

18.

Hanley S, Liu S, Lipsett M, Castellarin M, Rosenberg L, Tchervenkov J . Tumor necrosis factor-alpha production by human islets leads to postisolation cell death. Transplantation. 2006; 82(6):813-8. DOI: 10.1097/01.tp.0000234787.05789.23. View

19.

Gunji T, Saito T, Sato Y, Matsuyama S, Ise K, Kimura T . Mitomycin-C treatment followed by culture produces long-term survival of islet xenografts in a rat-to mouse model. Cell Transplant. 2008; 17(6):619-29. DOI: 10.3727/096368908786092720. View

20.

Noguchi H, Naziruddin B, Jackson A, Shimoda M, Ikemoto T, Fujita Y . Fresh islets are more effective for islet transplantation than cultured islets. Cell Transplant. 2012; 21(2-3):517-23. DOI: 10.3727/096368911X605439. View