Collagenase: Moving from black box to transparent knowledge
Bob McCarthy
Friday, January 22, 2016
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Chapters
Introduction
R01-Equivalent investigators Competing applicants, awardees, and funding rates
Crude collagenase: weak link in experimental chain to recover cells from tissue
Why continue to use these reagents?
Strengthening the weak link
Outline of the presentation
Mechanism of Tissue Dissociation
ECM designed to resist proteolysis
Proposed Mechanism of Action
Consistency: Crude & Enriched Collagenase
Raw material from minimally processed culture supernatant
Physical analysis of crude collagenase
Anion exchange chromatography analysis
Correlation of physical properties to collagen degradation (CDA) activity
Type 1 Worthington (WBC) Crude collagenase
Analysis of 3 lots enriched WBC Type I
Analysis of two lots of Sigma Type XI, enriched but "ill defined"
Comparison of effectiveness of different forms C1 for recovery porcine islets
Consistency: DE Collagenase
DE Collagenase product line
Analysis of DE Collagenase 100
Old vs New
Composition: Crude & Enriched Collagenase
Type defined by enzyme activities
How activities use to categorize Worthington Collagenase
VitaCyte Collagen degradation activity (CDA) assay functionally assesses collagenase
Breadth of composition in ill defined collagenase products
Compostion compared to purified products
Composition: DE Collagenase
Analysis of enzyme activities
Matrix provides guidance for use
Tip for optimizing composition
Convenience: DE Collagenase
Stable on storage
Transition to translational medicine
Take Home Message
Changing the paradigm
VitaCyte: a company committed to transforming tissue dissociation
Thank you
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