Multicenter Assessment of Animal-free Collagenase AF-1 for Human Islet Isolation

Abstract

Animal-free (AF) SERVA Collagenase AF-1 and Neutral Protease (NP) AF GMP Grade have recently become available for human islet isolation. This report describes the initial experiences of 3 different islet transplant centers. Thirty-four human pancreases were digested using 1 vial of the 6 different lots of Collagenase AF-1 (2,000–2,583 PZ-U/vial) supplemented with 4 different lots of NP AF in a range of 50 to 160 DMC-U per pancreas. Isolation, culture, and quality assessment were performed using standard techniques as previously described. All data are presented as mean ± standard error of the mean (SEM). Variability of pancreas weight was associated with a wide range of collagenase and NP activities, ranging from 12.7 to 46.6 PZ-U/g (26.0 ± 1.5 PZ-U/g) and 0.4 to 3.0 DMC-U/g (1.5 ± 0.1 DMC-U/g), respectively. Postpurification islet yield was 296,494 ± 33,620 islet equivalents (IEQ) equivalent to 3,274 ± 450 IEQ/g with a purity of 55.9% ± 3.2%. Quality assessment performed after 2 to 4 d of culture demonstrated a viability of 88.1% ± 1.5% and a stimulation index of 3.7 ± 0.7. Eighteen of the 34 preparations were transplanted into type 1 diabetic patients equivalent to a transplantation rate of 52.9%. Six preparations, which were infused into patients as first transplant, could be analyzed and increased the fasting C-peptide level from 0.11 ± 0.08 pretransplant to 1.23 ± 0.24 and 2.27 ± 0.31 ng/mL 3 and 6 mo posttransplant (P < 0.05), respectively. Insulin requirements were simultaneously reduced at the same time from 39.2 ± 3.8 IU/d before transplantation to 10.8 ± 4.1 and 4.0 ± 2.3 IU/d, after 3 and 6 mo posttransplant (P < 0.05), respectively. This study demonstrates the efficiency of AF SERVA Collagenase AF-1 and NP AF for clinical islet isolation and transplantation. The new plant-based production process makes these products a safe new option for the islet field.

Impact of ischemia time on islet isolation success and posttransplantation outcomes: A retrospective study of 452 pancreas isolations

“…We detected a significant increase in isolation success over the last years when comparing with the period between 2008 and 2013. This result is in accordance with a recent report of the Collaborative Islet and Transplantation Registry, 5 and is probably explained by improvements in the commercially available collagenase blends 16,23 . IGL‐1 has become the most commonly used preservation solution in recent years.…”

“…Toso et al reported that CIT >8 hours and ORT (secondary warm ischemia time) >30 minutes was associated with a reduced islet yield 10 . These two studies were published over 15 years ago, and significant advances in the development of novel enzyme blends may have contributed to mitigate the impact of parameters previously shown as deleterious 16,23 . In a more recent study, Berkova et al reported among other variables that a CIT >8 hours had significant adverse effects on isolation success 7 .…”

Reliable isolation of human mesenchymal stromal cells from bone marrow biopsy specimens in patients after allogeneic hematopoietic cell transplantation

“…Despite a higher yield, the collagenase-based MSC isolation technique from bone marrow biopsy specimens will not immediately replace the manufacturing of therapeutic MSC batches starting from bone marrow aspirates because most collagenase preperations are still poorly standardized [37] and bear the risk of transmiting infectious diseases [38].…”

Challenges, highlights, and opportunities in cellular transplantation: A white paper of the current landscape

“…Many consider the preparation required for cellular transplantation as a significant disadvantage, and the numerous steps of islet isolation are illustrative (Table 3). 10 Similarly, as described below, other cellular transplants will need continued basic and clinical research to achieve reproducible preparation for broad application 11,12 …”