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

Simona Campora, Giulio Ghersi, Silvia Saladino, Giulio Ghersi, Monica Salamone

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5 Citations (Scopus)

Abstract

Collagenases class I (Col G) and class II (Col H) currently available for tissue dissociation are producedfrom Clostridium histolyticum (human pathogen) strains. In the processes of extraction of the cells from thetissue, combined activity of both classes of enzymes is required. CI and CII are complementary indegrading collagen. ABIEL recently produced the collagenase class I and II using the recombinant DNAtechnologies (PCT WO 2011/073925 A9). The enzymes were produced in E. coli and purified by affinitychromatography. The method of production adopted allows absolute control of the final composition ofthese enzymes, as well as their stability, purity, activity, absence of toxicity and higher reproducibility ofbatches of collagenase. The two collagenases produced separately have been used in conjunctionaccording to precise proportions to dissociate calvaria and liver of the BALB/c mouse and bovine hoof. Theanalysis carried out on all isolated cell populations suggest that the cells maintain the structural andfunctional integrity of specific tissues/organs originating. Recombinant Col G and Col H enzymes representa promising tool for tissue dissociation.
Original languageEnglish
Pages247-252
Number of pages6
Publication statusPublished - 2014

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All Science Journal Classification (ASJC) codes

  • Chemical Engineering(all)

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