Improve islet yields and quality when clinical grade pancreata are preserved by the two-layer method
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Cell and Tissue Banking (2006) 7:195–201 DOI 10.1007/s10561-006-0002-0
Improved islet yield and quality from clinical grade pancreata preserved by the two-layer method Guangming Zhang1,5, Shinichi Matsumoto2, Helen Newman3, Wenjing Wang1,5, D. Michael Strong1, R. Paul Robertson4,5 and Jo-Anna Reems1,5,* 1
Northwest Tissue Center at the Puget Sound Blood Center, Seattle, WA 98104, USA; 2Kyoto University Hospital Transplant Unit, Kyoto, Japan; 3Veterinary Transplant Services, Inc., Kent, Washington, USA; 4 Pacific Northwest Research Institute, Seattle, Washington, USA; 5Department of Medicine, University of Washington, Seattle, Washington, USA; *Author for correspondence (e-mail: [email protected]; phone: +1-206-292-2317; fax: +1-206-343-1776) Received 13 April 2005; accepted in revised form 21 January 2006
Key words: Clinical grade pancreas, Islet transplant, Two-layer method
Abstract Background: Research grade pancreata preserved by the two-layer method (TLM) yield significantly greater numbers of islets than organs stored with University of Wisconsin solution (UW). The goal of this study was to determine whether this would hold true for pancreata that met selection criteria for clinical grade organs. Methods: Pancreata were chosen based upon a pre-defined set of criteria used for selecting clinical grade pancreata. Thirteen of these organs were preserved in UW and five pancreata were preserved by the TLM. Islets were isolated and evaluated according to the Edmonton protocol. Results: The average preservation time was significantly longer for organ preserved with TLM (9.5 + 2.0 h) as compared to UW (5.8 + 0.6 h, p = 0.015). The pancreata from the TLM group resulted in a significant increase in islet yields (3588 ± 500 vs. 2536 ± 312 IE/g pancreas, p < 0.05). Visual scoring of islets indicated that islets were better from the TLM group (8.3 ± 0.3 vs. 7.3 ± 0.2), and islet survival rates after culture were higher from organs stored with the TLM (87 ± 17 vs. 55 ± 7.4, p < 0.05). Other parameters such as viability, insulin content, and stimulation index were similar between the two groups. All the preparations from the TLM group, but only 54% of preparations from the UW group, qualified for islet transplantation. The two recipients receiving islets from the TLM group, experienced reduced daily insulin requirements and C-peptide levels increased. Conclusion: Compared to storage with UW, exposure of pancreata to the TLM resulted in greater islet yields and an improved quality of islets despite longer preservation. Consequently, pancreata that meet clinical grade status should be preserved by the TLM prior to islet isolation.
Introduction Islet transplantation reported by different transplant centers is highly effective for reversing the diabetic state of patients with type 1 diabetes (Shapiro et al. 2000, 2003; Ricordi 2003; Ricordi
et al. 2003). With the newer steroid-free, sirolimusbased immunosuppression protocols over 80% of patients have been able to discontinue insulin (Ricordi 2003; Shapiro et al. 2003). In general, is
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