Serva collagenase NB1: a new enzyme preparation for human islet isolation

Bucher, Pascal Alain Robert; Máthé, Z.; Bosco, Domenico; Andrès, Axel; Kurfuerst, M.; Rämsch-Günther, Nicole; Bühler, Léo H.; Morel, P.; Berney, Thierry

doi:10.1016/j.transproceed.2004.04.023

Cited by 19 publications

(15 citation statements)

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“…In 2007, NB-1 and NB enzymes (EC-A and EC-B) were the only available purified enzymes. Despite early reports describing the ability of EC-A and EC-B to produce similar islet yields to those achieved with L-HI (25, 26), many researchers found it difficult to reproduce these results. Previously, we reported that a higher proportion of intact-C1 with a molecular weight of 116 kDa (C1 116kDa ) was found in L-HI when compared to EC-A or EC-B (17).…”

Section: Discussionmentioning

confidence: 98%

A New Enzyme Mixture to Increase the Yield and Transplant Rate of Autologous and Allogeneic Human Islet Products

et al. 2012

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Background The optimal enzyme blend which maximizes human islets yield for transplantation remains to be determined. In this study, we evaluated 8 different enzyme combinations (ECs) in an attempt to improve islet yield. The ECs consisted of purified, intact, or truncated class 1 (C1) and class 2 (C2) collagenases from Clostridium histolyticum (Ch) as well as neutral protease (NP) from Bacillus thermoproteolyticus rokko (thermolysin) or Ch (ChNP). Methods We report the results of 249 human islet isolations, including 99 deceased donors (research n=57, clinical n=42) and 150 chronic pancreatitis pancreases. We prepared a new enzyme mixture (NEM) composed of intact C1 and C2 collagenases and ChNP instead of using thermolysin. The NEM was first tested in split pancreas (n=5) experiments and then used for islet autologous (n=21) and allogeneic transplantation (n=10). Islet isolation outcomes from 8 different Ecs were statistically compared using multivariate analysis. Results The NEM consistently achieved higher islet yields from pancreatitis (p<0.003) and deceased donor pancreases (p<0.001) than other standard ECs. Using the NEM, islet products met release criteria for transplantation from 8 of 10 consecutive pancreases, averaging 6510±2150 IEQ/g pancreas and 694,681±147,356 total IEQ/transplantation. In autologous isolation, the NEM yielded >200,000 IEQ from 19 of 21 pancreases (averaging 422,893±181,329 total IEQ and 5979±1469 IEQ/kg recipient body weight) regardless of the severity of fibrosis. Conclusions A new enzyme mixture composed of Clostridium histolyticum neutral protease with CIzyme high intact C1 collagenase recovers higher islet yield from deceased and pancreatitis pancreases while retaining islet quality and function.

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Section: Discussionmentioning

confidence: 98%

A New Enzyme Mixture to Increase the Yield and Transplant Rate of Autologous and Allogeneic Human Islet Products

et al. 2012

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“…10,36 Collagenase NB1 and NP are reconstituted separately and mixed before use to minimize degradation of the collagenase by NP. Consequently, the amounts of collagenase and NP can be adjusted independently to match the size of pancreas being processed.…”

Section: Discussionmentioning

confidence: 99%

“…Consequently, the amounts of collagenase and NP can be adjusted independently to match the size of pancreas being processed. 10,36 According to the certificate of analysis, the biochemical activity of Serva NB1 is similar to Liberase HI, but its parameters for optimal use are different and many experienced centers have had inconsistent results using this product. 10,36 Interestingly, recent data from CITR show improvement in primary efficacy and safety outcomes of islet transplantation in recipients who received transplants in 2007-2010 compared with those in 1999-2006, with fewer islet infusions and adverse events per recipient.…”

Section: Discussionmentioning

confidence: 99%

“…10,36 According to the certificate of analysis, the biochemical activity of Serva NB1 is similar to Liberase HI, but its parameters for optimal use are different and many experienced centers have had inconsistent results using this product. 10,36 Interestingly, recent data from CITR show improvement in primary efficacy and safety outcomes of islet transplantation in recipients who received transplants in 2007-2010 compared with those in 1999-2006, with fewer islet infusions and adverse events per recipient. 3 Although these dates coincide with the switch in the use of Liberase HI enzyme to Serva NB1 enzyme, it is not possible to conclude that the improvement of the islet transplantation results in more recent years is only due to the utilization of Serva NB1 blend, since no randomized clinical trials were performed specifically to compare these 2 enzymes.…”

Section: Discussionmentioning

confidence: 99%

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Different digestion enzymes used for human pancreatic islet isolation: A mixed treatment comparison (MTC) meta-analysis

Rheinheimer

Ziegelmann

Carlessi

et al. 2014

Islets

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Daisy Crispim (2014) Different digestion enzymes used for human pancreatic islet isolation: A mixed treatment comparison (MTC) meta-analysis, Islets, 6:4, e977118, DOI: 10.4161/19382014.2014.977118 To link to this article: https://doi.org/10. 4161/19382014.2014 Collagenases are critical reagents determining yield and quality of isolated human pancreatic islets and may affect islet transplantation outcome. Some islet transplantation centers have compared 2 or more collagenase blends; however, the results regarding differences in quantity and quality of islets are conflicting. Thus, for the first time, a mixed treatment comparison (MTC) meta-analysis was carried out to compile data about the effect of different collagenases used for human pancreas digestion on islet yield, purity, viability and stimulation index (SI). Pubmed, Embase and Cochrane libraries were searched. Of 755 articles retrieved, a total of 15 articles fulfilled the eligibility criteria and were included in the MTC meta-analysis. Our results revealed that Vitacyte and Liberase MTF were associated with a small increase in islet yield (islet equivalent number/g pancreas) when compared with Sevac enzyme [standardized mean difference (95% credible interval -CrI) D ¡2.19 (¡4.25 to ¡0.21) and ¡2.28 (¡4.49 to ¡0.23), respectively]. However, all other enzyme comparisons did not show any significant difference regarding islet yield. Purity and viability percentages were not significantly different among any of the analyzed digestion enzymes. Interestingly, Vitacyte and Serva NB1 were associated with increased SI when compared with Liberase MTF enzyme [unstandardized weighted mean difference (95% CrI) D ¡1.69 (¡2.87 to ¡0.51) and ¡1.07 (¡1.79 to ¡0.39), respectively]. In conclusion, our MTC meta-analysis suggests that the digestion enzymes currently being used for islet isolation works with similar efficiency regarding islet yield, purity and viability; however, Vitacyte and Serva NB1 enzymes seem to be associated with an improved SI as compared with Liberase MTF.

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“…Several groups have reported the comparison between Liberase HI and collagenase NB1 blends in terms of the efficiency of islet isolations yield and islet quality (2, 8, 9, 10, 39). Recently Brandhorst et al .…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Viable β-Cell Mass is Useful for Selecting Collagenase for Human Islet Isolation: Comparison of Collagenase NB1 and Liberase HI

et al. 2012

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Objectives The selection of enzyme blend is critical for the success of human islet isolations. Liberase HI collagenase (Roche) has been introduced in the 1990’s and widely used for clinical islet transplantation. More recently, a blend collagenase NB1 has been rendered available. The aim of this study was to evaluate the isolation outcomes and islet quality comparing human islet cells processed using NB1 and Liberase HI. Methods A total of 90 isolations processed using NB1 (n=40) or Liberase HI (n=50) was retrospectively analyzed. Islet yield, function in vitro and in vivo, cellular (including β-cell specific) viability and content, as well as isolation related factors were compared. Results No significant differences in donor related factors were found between the groups. There were also no significant differences in islet yields (NB1 vs. Liberase; 263,389±21,550 vs. 324,256±27,192 IEQ; P = n.s., respectively). The pancreata processed with NB1 showed a significantly longer digestion time (18.6±0.7 vs. 14.5±0.5 min, P <0.01), lower β-cell viability (54.3±3.4 vs. 72.0±2.1%, P < 0.01), β-cell mass (93,671±11,150 vs. 148,961± 12,812βIEQ, P<0.01) and viable β-cell mass (47,317±6,486 vs. 106,631±10,228 VβIEQ, P < 0.01) than Liberase HI. In addition, islets obtained with Liberase showed significantly better graft function in in vivo assessment of islet potency. Conclusions The utilization of collagenase NB1 in human islet isolation was associated with significantly lower β-cell viability, mass and islet potency in vivo in our series when compared to Liberase HI even though there was no significant difference in islet yields between the groups. Evaluation of viable β-cell mass contained in human islet preparations will be useful for selecting enzyme blends.

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Serva collagenase NB1: a new enzyme preparation for human islet isolation

Cited by 19 publications

References 4 publications

A New Enzyme Mixture to Increase the Yield and Transplant Rate of Autologous and Allogeneic Human Islet Products

A New Enzyme Mixture to Increase the Yield and Transplant Rate of Autologous and Allogeneic Human Islet Products

Different digestion enzymes used for human pancreatic islet isolation: A mixed treatment comparison (MTC) meta-analysis

Evaluation of Viable β-Cell Mass is Useful for Selecting Collagenase for Human Islet Isolation: Comparison of Collagenase NB1 and Liberase HI

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