Clostripain, the Missing Link in the Enzyme Blend for Efficient Human Islet Isolation

Ståhle, Magnus; Foss, Aksel; Gustafsson, Bengt; Lempinen, Marko; Lundgren, Torbjörn; Rafael, Ehab; Tufveson, Gunnar; Korsgren, Olle; Friberg, Andrew

doi:10.1097/txd.0000000000000528

Cited by 18 publications

(24 citation statements)

References 45 publications

(40 reference statements)

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“…As the patients usually require a repeated islet infusion, and not all islet isolations lead to clinical transplantation, the real cost for islet transplantation is higher. The islet isolation process does therefore represent a significant potential for cost reduction, and investigations into increasing the success rate and developing more efficient, low‐cost isolation techniques are ongoing . It is also important to note that isolations that fail to yield transplantation level amount of islets often result in human islets available to research .…”

Section: Discussionmentioning

confidence: 99%

“…The islet isolation process does therefore represent a significant potential for cost reduction, and investigations into increasing the success rate and developing more efficient, low-cost isolation techniques are ongoing. 25,26 It is also important to note that isolations that fail to yield transplantation level amount of islets often result in human islets available to research. 27,28 This is a favorable part of an islet program that should not be ignored.…”

Section: Discussionmentioning

confidence: 99%

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Cost and clinical outcome of islet transplantation in Norway 2010‐2015

Schive

Foss

Sahraoui

et al. 2016

Clinical Transplantation

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Islet transplantation is a minimally invasive β-cell replacement strategy. Islet transplantation is a reimbursed treatment in Norway. Here, we summarize the cost and clinical outcome of 31 islet transplantations performed at Oslo University Hospital (OUS) from January 2010 to June 2015. Patients were retrospectively divided into three groups. Thirteen patients received either one or two islet transplantation alone (ITA), while five patients received islet transplantation after previous solid organ transplantation. For the group receiving 2 ITA, Kaplan-Meier estimates show an insulin independence of 20% more than 4 years after their last transplantation. An estimated 70% maintain at least partial graft function, defined as fasting C-peptide >0.1 nmol L , and 47% maintain a HbA1c below 6.5% or 2 percent points lower than before ITA. For all groups combined, we estimate that 44% of the patients have a 50% reduction in insulin requirement 4 years after the initial islet transplantation. The average cost for an islet transplantation procedure was 347 297±60 588 NOK, or 35 424±6182 EUR, of which isolation expenses represent 34%. We hereby add to the common pool of growing experience with islet transplantation and also describe the cost of the treatment at our center.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Cost and clinical outcome of islet transplantation in Norway 2010‐2015

Schive

Foss

Sahraoui

et al. 2016

Clinical Transplantation

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“…Moreover, a recent study has shown that supplementation of CIzyme Collagenase, Thermolysin with Clostripain improved human islet isolation outcomes. 45 The new Liberase MTF C/T is a highly purified GMP-grade and animal tissue free product with lower endotoxin levels. Therefore, lot-to-lot variability has been minimum providing very stable enzyme mixtures.…”

Section: Discussionmentioning

confidence: 99%

The Choice of Enzyme for Human Pancreas Digestion Is a Critical Factor for Increasing the Success of Islet Isolation

Valiente

McFadden

et al. 2015

Transplantation Direct

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Background We evaluated three commercially available enzymes for pancreatic digestion by comparing key parameters during the islet isolation process, as well as islet quality post-isolation. Methods Retrospectively compared and analyzed islet isolations from pancreata using three different enzyme groups: Liberase HI (n=63), Collagenase NB1/Neutral Protease (NP) (n=43), and Liberase Mammalian Tissue Free Collagenase/Thermolysin (MTF C/T) (n=115). A standardized islet isolation and purification method was used. Islet quality assessment was carried out using islet count, viability, in vitro glucose-stimulated insulin secretion (GSIS), glucose-stimulated oxygen consumption rate (ΔOCR), and in vivo transplantation model in mice. Results Donor characteristics were not significantly different among the three enzyme groups used in terms of age, sex, hospital stay duration, cause of death, body mass index (BMI), hemoglobin A1c (HbA1c), cold ischemia time (CIT), and pancreas weight. Digestion efficacy (percentage of digested tissue by weight) was significantly higher in the Liberase MTF C/T group (73.5 ± 1.5 %) when compared to the Liberase HI group (63.6 ± 2.3 %) (p<0.001) and the Collagenase NB1/NP group (61.7 ± 2.9%) (p<0.001). The stimulation index for GSIS was significantly higher in the Liberase MTF C/T group (5.3 ± 0.5) as compared to the Liberase HI (2.9 ± 0.2) (p<0.0001) and the Collagenase NB1/NP (3.6 ± 2.9) (p=0.012) groups. Furthermore, the Liberase MTF C/T enzymes showed the highest success rate of transplantation in diabetic NOD Scid mice (65%), which was significantly higher than the Liberase HI (42%, p=0.001) and the Collagenase NB1/NP enzymes (41%, p<0.001). Conclusions Liberase MTF C/T is superior to Liberase HI and Collagenase NB1/NP in terms of digestion efficacy and glucose-stimulated insulin secretion in vitro. Moreover, Liberase MTF C/T had a significantly higher success rate of transplantation in diabetic NOD Scid mice compared to Liberase HI and Collagenase NB1/NP enzymes.

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“…Although the NB-1 Collagenase and NB Protease continues to be used by many laboratories, one group found that those lots with higher amounts of clostripain activity gave higher human islet yields [49]. Another group found the addition of clostripain to Collagenase HA/thermolysin enzyme mixtures increased islet yields [50]. The relevance of adding clostripain to purified collagenase-protease enzyme mixtures will be discussed in more detail below.…”

Section: ++++mentioning

confidence: 99%

“…Many laboratories are unaware of this change and continue to assume that thermolysin in the MTF kit is at a similar concentration as Liberase HI (version 4). Others addressed the problem by reducing the amount of thermolysin used in the isolation procedure [6,50].…”

Section: Selection and Dose Of Neutral Protease Is Critical For Succementioning

confidence: 99%

Evolution of Enzyme Requirements for Human Islet Isolation

McCarthy¹,

Green²,

Dwulet³

2018

obm transplant

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Islet transplantation is becoming an established treatment option for managing a subset of adult patients who have type 1 diabetes mellitus. The success of this procedure is dependent upon the recovery of a sufficient number of functional human islets from donor organs for subsequent transplant. Here, the use of optimized bacterial collagenase-neutral protease enzyme mixtures has been shown to affect the yield and quality (defined by viability and glucose-stimulated insulin secretion) of islets recovered from human pancreata. However, few reports provide a systematic approach to correlate the biochemical characteristics or dose of collagenases and proteases to the recovery of functional islets. The focus of this review is to close this gap. The review summarizes five key advances toward an understanding of how Clostridium histolyticum collagenase and bacterial neutral proteases support the release of islets from human pancreata and key collagenase biochemical characteristics that lead to higher human islet yields. This information provides a foundation to develop a model of enzymatic tissue dissociation that can serve as a guide in assessing the effectiveness of collagenase or neutral protease enzymes used for human islet isolation. One key conclusion is the importance of excess amounts of collagen degradation activity in the islet isolation procedure to ensure

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Clostripain, the Missing Link in the Enzyme Blend for Efficient Human Islet Isolation

Abstract: Supplemental digital content is available in the text.

Cited by 18 publications

References 45 publications

Cost and clinical outcome of islet transplantation in Norway 2010‐2015

Cost and clinical outcome of islet transplantation in Norway 2010‐2015

The Choice of Enzyme for Human Pancreas Digestion Is a Critical Factor for Increasing the Success of Islet Isolation

Evolution of Enzyme Requirements for Human Islet Isolation

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