Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and II

Collagenases class I (Col G) and class II (Col H) currently available for tissue dissociation are produced from Clostridium histolyticum (human pathogen) strains. In the processes of extraction of the cells from the tissue, combined activity of both classes of enzymes is required. CI and CII are com...

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Main Authors: M. Salamone, S. Saladino, M. Pampalone, S. Campora, G. Ghersi
Format: Article
Language:English
Published: AIDIC Servizi S.r.l. 2014-09-01
Series:Chemical Engineering Transactions
Online Access:https://www.cetjournal.it/index.php/cet/article/view/5647
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author M. Salamone
S. Saladino
M. Pampalone
S. Campora
G. Ghersi
author_facet M. Salamone
S. Saladino
M. Pampalone
S. Campora
G. Ghersi
author_sort M. Salamone
collection DOAJ
description Collagenases class I (Col G) and class II (Col H) currently available for tissue dissociation are produced from Clostridium histolyticum (human pathogen) strains. In the processes of extraction of the cells from the tissue, combined activity of both classes of enzymes is required. CI and CII are complementary in degrading collagen. ABIEL recently produced the collagenase class I and II using the recombinant DNA technologies (PCT WO 2011/073925 A9). The enzymes were produced in E. coli and purified by affinity chromatography. The method of production adopted allows absolute control of the final composition of these enzymes, as well as their stability, purity, activity, absence of toxicity and higher reproducibility of batches of collagenase. The two collagenases produced separately have been used in conjunction according to precise proportions to dissociate calvaria and liver of the BALB/c mouse and bovine hoof. The analysis carried out on all isolated cell populations suggest that the cells maintain the structural and functional integrity of specific tissues/organs originating. Recombinant Col G and Col H enzymes represent a promising tool for tissue dissociation.
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spelling doaj.art-4ee571b17da84af1ada061ecc98473b92022-12-21T17:13:48ZengAIDIC Servizi S.r.l.Chemical Engineering Transactions2283-92162014-09-013810.3303/CET1438042Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and IIM. SalamoneS. SaladinoM. PampaloneS. CamporaG. GhersiCollagenases class I (Col G) and class II (Col H) currently available for tissue dissociation are produced from Clostridium histolyticum (human pathogen) strains. In the processes of extraction of the cells from the tissue, combined activity of both classes of enzymes is required. CI and CII are complementary in degrading collagen. ABIEL recently produced the collagenase class I and II using the recombinant DNA technologies (PCT WO 2011/073925 A9). The enzymes were produced in E. coli and purified by affinity chromatography. The method of production adopted allows absolute control of the final composition of these enzymes, as well as their stability, purity, activity, absence of toxicity and higher reproducibility of batches of collagenase. The two collagenases produced separately have been used in conjunction according to precise proportions to dissociate calvaria and liver of the BALB/c mouse and bovine hoof. The analysis carried out on all isolated cell populations suggest that the cells maintain the structural and functional integrity of specific tissues/organs originating. Recombinant Col G and Col H enzymes represent a promising tool for tissue dissociation.https://www.cetjournal.it/index.php/cet/article/view/5647
spellingShingle M. Salamone
S. Saladino
M. Pampalone
S. Campora
G. Ghersi
Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and II
Chemical Engineering Transactions
title Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and II
title_full Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and II
title_fullStr Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and II
title_full_unstemmed Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and II
title_short Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and II
title_sort tissue dissociation and primary cells isolation using recombinant collagenases class i and ii
url https://www.cetjournal.it/index.php/cet/article/view/5647
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