Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet Isolation

Shima, Hiroki; Inagaki, Akiko; Imura, Takehiro; Yamagata, Yuriko; Watanabe, K.; Igarashi, Kazuhiko; Goto, Masafumi; Murayama, Kazutaka

doi:10.1155/2016/4396756

Cited by 15 publications

(15 citation statements)

References 36 publications

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“…Several studies have also shown detrimental effects of enzymatic islet isolation on peripheral islet ECM, such as on collagens [ 2 , 6 ] and laminins [ 119 ]. Collagenases digest several collagen types, such as types I, III, IV and V [ 25 ]. This has a dramatic impact on cell viability [ 27 ].…”

Section: Considerations For Engineering and Future Applications Of Ecmentioning

confidence: 99%

“…Although selective, the process of enzymatic degradation of exocrine connections is not specific [ 2 ] and, as a consequence, many ECM components that surround the islets and interconnect endocrine cells are also damaged [ 2 , 22 ], affecting islet function [ 6 , 21 , 23 , 24 ]. After isolation using ECM-degrading collagenases, the whole microvasculature of the islet is destroyed [ 25 ] and islet cells undergo cell-death processes, such as anoikis, necroptosis and necrosis [ 2 , 22 , 26 , 27 ]. To amplify matters, these processes are associated with the release of highly inflammatory danger-associated molecular patterns (DAMPs) that contribute to immune responses against pancreatic islets [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

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Extracellular matrix molecules and their potential contribution to the function of transplanted pancreatic islets

2018

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Extracellular matrix (ECM) molecules are responsible for structural and biochemical support, as well as for regulation of molecular signalling and tissue repair in many organ structures, including the pancreas. In pancreatic islets, collagen type IVand VI, and laminins are the most abundant molecules, but other ECM molecules are also present. The ECM interacts with specific combinations of integrin α/β heterodimers on islet cells and guides many cellular processes. More specifically, some ECM molecules are involved in beta cell survival, function and insulin production, while others can fine tune the susceptibility of islet cells to cytokines. Further, some ECM induce release of growth factors to facilitate tissue repair. During enzymatic isolation of islets for transplantation, the ECM is damaged, impacting islet function. However, restoration of the ECM in human islets (for example by adding ECM to the interior of immunoprotective capsules) has been shown to enhance islet function. Here, we provide current insight into the role of ECM molecules in islet function and discuss the clinical potential of ECM manipulation to enhance pancreatic islet function and survival.

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Section: Considerations For Engineering and Future Applications Of Ecmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Extracellular matrix molecules and their potential contribution to the function of transplanted pancreatic islets

2018

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“…This finding is particularly remarkable since collagen-VI is resistant toward the activity of bacterial collagenases [136,137], suggesting a distinct protective morphological wall during enzyme-mediated pancreas digestion. Moreover, collagen-V, the second most strongly expressed collagen subtype in the human peri-islet basement membrane, can be degraded by CC-I, but not by CC-II [138]. This finding is in clear contradiction to the proposition that CC-II is the most relevant isoform for islet release when compared with CC-I [47,59].…”

Section: Extracellular Matrix Degradationmentioning

confidence: 65%

Enzyme Development for Human Islet Isolation: Five Decades of Progress or Stagnation?

Brandhorst¹,

Brandhorst²,

Prv.³

2017

Rev Diabet Stud

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In comparison to procedures used for the separation of individual cell types from other organs, the process of human pancreatic islet isolation aims to digest the pancreatic exocrine matrix completely without dispersing the individual cells within the endocrine cell cluster. This objective is unique within the field of tissue separation, and outlines the challenge of islet isolation to balance two opposing priorities. Although significant progress has been made in the characterization and production of enzyme blends for islet isolation, there are still numerous areas which require improvement. The ultimate goal of enzyme production, namely the routine production of a consistent and standardized enzyme blend, has still not been realized. This seems to be mainly the result of a lack of detailed knowledge regarding the structure of the pancreatic extracellular matrix and the synergistic interplay between collagenase and different supplementary proteases during the degradation of the extracellular matrix. Furthermore, the activation of intrinsic proteolytic enzymes produced by the pancreatic acinar cells, also impacts on the chance of a successful outcome of human islet isolation. This overview discusses the challenges of pancreatic enzymatic digestion during human islet isolation, and outlines the developments in this field over the past 5 decades.

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“…1 ). Collagenase is also used in islet isolation from human pancreas for transplantation surgery of type 1 diabetes [24] , [37] , [38] , [39] , [40] , [41] , [42] , [43] . It was recently reported that proteases with trypsin-like activity strongly influence islet isolation from the human pancreas [23] .…”

Section: Discussionmentioning

confidence: 99%

Technical advantage of recombinant collagenase for isolation of muscle stem cells

Ishii

Suzuki

Mabuchi

et al. 2017

Regenerative Therapy

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BackgroundMuscle satellite cells are resident skeletal muscle stem cells responsible for muscle regeneration. Isolation of satellite cells is a critical process for clinical application such as drug screening and cell transplantation. Fluorescence-activated cell sorting (FACS) enables the direct isolation of satellite cells from muscle tissue. During the process used to isolate satellite cells from skeletal muscle, enzymatic digestion is the first step. Therefore, the evaluation and standardization of enzymes is important not only for reproducibility of cellular yield and viability, but also for traceability of material used in protocols.MethodsThe comparison of muscle digestion was performed either by a mixture of recombinant collagenase G (ColG) and collagenase H (ColH) or by a conventional collagenase II. The degree of cell damage and surface antigen expression upon collagenase treatment were analyzed by FACS. To investigate whether satellite cells isolated using recombinant collagenase can regenerate injured muscle, satellite cells were cultured, transplanted into injured muscles, and analyzed by immunostaining.ResultsWe show that ColG and ColH were efficient to isolate satellite cells from mouse skeletal muscle tissue. Digestion with a combination of ColG and ColH enriched satellite cells with intact surface antigens such as α7 and β1 integrins. Furthermore, satellite cells isolated using ColG and ColH dramatically proliferated and remained undifferentiated in vitro. When transplanted, satellite cells isolated using ColG and ColH enhanced the therapeutic efficacy in vivo.ConclusionsOur results provide an efficient method of satellite cell preparation using recombinant collagenases with a high cell yield, viability of cells, and regeneration potency to fit the biological raw material criteria.

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Collagen V Is a Potential Substrate for Clostridial Collagenase G in Pancreatic Islet Isolation

Cited by 15 publications

References 36 publications

Extracellular matrix molecules and their potential contribution to the function of transplanted pancreatic islets

Extracellular matrix molecules and their potential contribution to the function of transplanted pancreatic islets

Enzyme Development for Human Islet Isolation: Five Decades of Progress or Stagnation?

Technical advantage of recombinant collagenase for isolation of muscle stem cells

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