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

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

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)963689720947098
JournalCell transplantation
Volume29
DOIs
StatePublished - Jan 1 2020

Keywords

  • diabetes
  • islet transplantation

ASJC Scopus subject areas

  • Biomedical Engineering
  • Cell Biology
  • Transplantation

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    Gołębiewska, J. E., Gołąb, K., Gorycki, T., Śledziński, M., Gulczyński, J., Żygowska, I., Wolnik, B., Hoffmann, M., Witkowski, P., Ricordi, C., Szurowska, E., Śledziński, Z., & Dębska-Ślizień, A. (2020). "Old School" Islet Purification Based on the Unit Gravity Sedimentation as a Rescue Technique for Intraportal Islet Transplantation-A Case Report. Cell transplantation, 29, 963689720947098. https://doi.org/10.1177/0963689720947098