'Old School' Islet Purification Based on the Unit Gravity Sedimentation as a Rescue Technique for Intraportal Islet Transplantation—A Case Report

Autor: Justyna E. Gołębiewska, Karolina Gołąb, Tomasz Gorycki, Maciej Śledziński, Jacek Gulczyński, Iwona Żygowska, Bogumił Wolnik, Michał Hoffmann, Piotr Witkowski, Camillo Ricordi, Edyta Szurowska, Zbigniew Śledziński, Alicja Dębska-Ślizień
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Zdroj: Cell Transplantation, Vol 29 (2020)
Druh dokumentu: article
ISSN: 1555-3892
09636897
DOI: 10.1177/0963689720947098
Popis: Here, we present a case that required a supplemental “old school” islet purification for a safe intraportal infusion. Following pancreas procurement from a brain-dead 26-year-old male donor (body mass index: 21.9), 24.6 ml of islet tissue was isolated after continuous density gradient centrifugation. The islet yield was 504,000 islet equivalent (IEQ), distributed among the following three fractions: 64,161 IEQ in 0.6 ml of pellet, 182,058 IEQ in 10 ml, and 258,010 IEQ in 14 ml with 95%, 20%, and 10% purity, respectively. After a 23-h culture, we applied supplemental islet purification, based on the separation of tissue subfractions during unit gravity sedimentation, a technique developed over 60 years ago (“old school”). This method enabled the reduction of the total pellet volume to 11.6 ml, while retaining 374,940 IEQ with a viability of over 90%. The final islet product was prepared in three infusion bags, containing 130,926 IEQ in 2.6 ml of pellet, 108,079 IEQ in 4 ml of pellet, and 135,935 IEQ in 5 ml of pellet with 65%, 40%, and 30% purity, respectively, and with the addition of unfractionated heparin (70 units/kg body weight). Upon the islet infusion from all three bags, portal pressure increased from 7 to 16 mmHg. Antithrombotic prophylaxis with heparin was continued for 48 h after the infusion, with target activated partial thromboplastin time 50–60 s, followed by fractionated heparin subcutaneous injections for 2 weeks. β-Cell graft function assessed on day 75 post-transplantation was good, according to Igls criteria, with complete elimination of severe hypoglycemic episodes and 50% reduction in insulin requirements. Time spent within the target glucose range (70–180 mg/dl) improved from 42% to 98% and HbA1c declined from 8.7% to 6.7%. Supplemental “old school” islet purification allowed for the safe and successful utilization of a robust and high-quality islet preparation, which otherwise would have been discarded.
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