Degraded Collagenase Deteriorates Islet Viability

Brandhorst, Heide; Raemsch-Guenther, N.; Raemsch, C.; Friedrich, O; Kurfuerst, M.; Korsgren, Olle; Brandhorst, Daniel

doi:10.1016/j.transproceed.2008.01.013

Cited by 9 publications

(8 citation statements)

References 5 publications

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“…8 More importantly, in the latter study, collagenase Ib was shown to reduce islet yields, and a correlation between degraded collagenase and reduced islet viability has been previously demonstrated. 3 In line with these findings, we 11 mM glucose, 0.15% NaHCO 3 (Invitrogen GmbH, Karlsruhe, Germany) and incubated at 37°C and 5% CO 2 . All functional experiments were performed after overnight culture.…”

Section: Discussionmentioning

confidence: 73%

“…1 Crucial criteria for obtaining maximum numbers of viable and functionally intact islets include collagenase composition and purity, and the duration of the enzymatic digestion of the pancreas. [2][3][4] Collagenases I and II, purified from Clostridium histolyticum, exhibit differences in their amino acid sequences and secondary structures, but show very similar catalytic domains. Collagenase I attacks residues near the ends of triple helical collagen domains, whereas collagenase II initially digests at more interior sites.…”

Section: Introductionmentioning

confidence: 99%

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A new collagenase blend increases the number of islets isolated from mouse pancreas

Yeşil¹,

Michel²,

Chwalek³

et al. 2009

Islets

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Section: Discussionmentioning

confidence: 73%

Section: Introductionmentioning

confidence: 99%

A new collagenase blend increases the number of islets isolated from mouse pancreas

Yeşil¹,

Michel²,

Chwalek³

et al. 2009

Islets

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“…Langerhans islets were isolated from healthy Wistar rats by a collagenase digestion method [35]. Wistar rats, bred at the University Medical Center, Mainz, 12–16 weeks old were used in all experiments.…”

Section: Methodsmentioning

confidence: 99%

Biocompatible Coating of Encapsulated Cells Using Ionotropic Gelation

et al. 2013

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The technique of immunoisolated transplantation has seen in the last twenty years improvements in biocompatibility, long term stability and methods for avoidance of fibrosis in alginate capsules. However, two major problems are not yet solved: living cellular material that is not centered in the capsule is not properly protected from the hosts’ immune system and the total transplant volume needs to be reduced. To solve these problems, we present a method for applying fully biocompatible alginate multilayers to a barium-alginate core without the use of polycations. We report on the factors that influence layer formation and stability and can therefore provide data for full adjustability of the additional layer. Although known for yeast and plant cells, this technique has not previously been demonstrated with mammalian cells or ultra-high viscous alginates. Viability of murine insulinoma cells was investigated by live-dead staining and live cell imaging, for murine Langerhans’ islets viability and insulin secretion have been measured. No hampering effects of the second alginate layer were found. This multi-layer technique therefore has great potential for clinical and in vitro use and is likely to be central in alginate matrix based immunoisolated cell therapy.

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“…A significant problem is that collagenase I is the most unstable component of Liberase, as the Ia form is rapidly autocatalytically degraded to the Ib form. Degraded collagenases have an adverse effect on islet viability [14]. There is a wide range of endotoxin contamination of traditional collagenase preparations compared with the endotoxin level of Liberase.…”

Section: Introductionmentioning

confidence: 99%

Matrix metalloproteinases as reagents for cell isolation

Knapinska

Amar

et al. 2016

Enzyme and Microbial Technology

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Cell isolation methods for therapeutic purposes have seen little advancement over the years. The original methods of stem cell and islet isolation using bacterial collagenases were developed in the early 1980s and are still used today. Bacterial collagenases are subject to autodegradation, and isolates obtained with these enzymes may be contaminated with endotoxins, reducing cell viability and contributing to toxicity in downstream applications. Here we describe a novel method for isolation of mesenchymal stem cells from adipose tissue (ADSC) utilizing recombinantly produced matrix metalloproteases (MMPs). The ADSCs isolated by MMPs displayed essentially identical morphological and phenotypical characteristics to cells isolated by bacterially-derived collagenase I and Liberase™. Samples isolated with MMPs and Liberase™ had comparable levels of CD73, CD90, and CD105. The adipogenic and osteogenic potential of the ADSCs isolated by MMPs was retained as compared to cells isolated with Liberase™. However, ADSCs isolated by Liberase™ displayed 6% contamination with other cells as per negative markers revealed by PE staining, as opposed to <1% for all MMP-treated samples. MMP-based cell isolation may contribute to optimization of transplantation technology.

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Degraded Collagenase Deteriorates Islet Viability

Cited by 9 publications

References 5 publications

A new collagenase blend increases the number of islets isolated from mouse pancreas

A new collagenase blend increases the number of islets isolated from mouse pancreas

Biocompatible Coating of Encapsulated Cells Using Ionotropic Gelation

Matrix metalloproteinases as reagents for cell isolation

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