Deterioration and variability of highly purified collagenase blends used in clinical islet isolation

Toshiyuki Yamamoto, Camillo Ricordi, Shari Messinger, Yasunaru Sakuma, Atsushi Miki, Rayner Rodriguez, Ross Haertter, Aisha Khan, Rodolfo Alejandro, Hirohito Ichii

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

BACKGROUND. The quality and stability of enzyme blends used in islet cell processing are critical for successful human islet isolation. A wide variability in enzymatic activity among lots of Liberase HI has been reported. This study examines the interlot and intralot variability of Liberase HI and the over-time deterioration of enzyme quality based on the analysis of islet isolation outcomes. METHODS. The data of 169 human isolations processed for clinical islet transplantation, using five different lots of Liberase HI, were retrospectively analyzed. Inter- and intralot variables in the islet isolation were assessed over a 15-month period. RESULTS. The analysis revealed significant interlot differences in the digestion time, prepurification islet counts, percent recovery, viability, and glucose stimulation insulin index. Moreover, a significant decrease in the pre- and postpurification islet yield per pancreas weight (IEQ/g) in isolations processed by two different enzyme lots used over a 15-month period was observed, suggesting a progressive deterioration of enzyme quality. CONCLUSIONS. Our data demonstrate a significant lot-to-lot related variability in islet isolation outcomes. In addition, the over-time decline in isolation outcomes processed using a single enzyme lot was observed even when the enzyme blends were used within the expiration dating specified by the manufacturer.

Original languageEnglish (US)
Pages (from-to)997-1002
Number of pages6
JournalTransplantation
Volume84
Issue number8
DOIs
StatePublished - Oct 1 2007

Keywords

  • Enzyme
  • Islet
  • Isolation
  • Liberase HI
  • Transplantation
  • Variables

ASJC Scopus subject areas

  • Transplantation
  • Immunology

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