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.

Salamone, M., Saladino, S., Pampalone, M., Campora, S., Ghersi, G. (2014). Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and II. In Chemical Engineeniring Transaction VOL. 38, 2014 (pp.247-252) [10.3303/CET1438042].

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

SALADINO, Silvia;Pampalone, M;CAMPORA, Simona;GHERSI, Giulio
2014-01-01

Abstract

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.
2014
IBIC 2014 4th INTERNATIONAL CONFERENCE ON INDUSTRIAL BIOTECHNOLOGY
Roma
8-11 Giugno, 2014
2014
6
Salamone, M., Saladino, S., Pampalone, M., Campora, S., Ghersi, G. (2014). Tissue Dissociation and Primary Cells Isolation Using Recombinant Collagenases Class I and II. In Chemical Engineeniring Transaction VOL. 38, 2014 (pp.247-252) [10.3303/CET1438042].
Proceedings (atti dei congressi)
Salamone, M; Saladino, S; Pampalone, M; Campora, S; Ghersi, G
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10447/99905
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