Overcoming the Limitations of Traditional Collagenase Products

Traditional collagenase products (i.e., crude or enriched collagenase) have been used for over 50 years to isolate cells from tissue. Little has changed in the selection and use of these products since the original introduction of crude collagenase in the early 1960’s for use in cell isolation procedures. Many laboratories pre-qualify lots prior to purchase

DE Collagenase Optimization Kit: A Fresh Approach to Defining Enzyme Composition and Dose for Maximal Cell Recovery

Currently, there is no simple method for defining optimal collagenase enzyme formulations for recovery of primary cells from tissue, or adherent cells from in vitro culture substrates. Crude or enriched collagenase products commonly used for these applications are minimally purified, ill-defined enzyme mixtures in which the collagenases and neutral proteases responsible for releasing cells from

Which One Are You Using?

Crude Collagenase vs. DE Collagenase   Crude Collagenase DE Collagenase Price $ to $$ $ to $$ % of Collagenase+ Protease 3-8% ≈ 74-85% Protease Activity Variable Fixed Evaluation New Lots EvaluatedBefore Use New Lot Screening Eliminated After Initial Evaluation Ease of Sterile Filtration Variable Easy Performance & Confidence ??? Variable Enzyme Activity Between Lots

Tired of Reliving Groundhog Day?

Do you feel like weatherman Phil Connors reliving Groundhog Day when you qualify new lots of traditional collagenase to isolate rodent islets? Avoid living in a time loop, break away from the need to qualify new lots of collagenase by switching to CIzyme RI, a purified, defined enzyme mixture designed for rodent islet isolation. At

New Paradigm for Collagenase Products

Essential Information on Enzyme Activities Required for Effective Cell Isolation Part A: Overview of Current Knowledge and State of the Art Overview Worthington Biochemicals began supplying Clostridium histolyticum collagenase for cell isolation in the early 1960s. Worthington’s Tissue Dissociation Guide summarizes the use of these enzymes for cell isolation. They categorized their collagenase products into

VitaCyte to Exhibit at the 9th EPITA Symposium/38th AIDPIT Workshop

VitaCyte and its European distributor, PeloBiotech, will exhibit at the Workshop held 27 to 29 January 2019 at Congresspark Igls-Innsbruck, Austria. VitaCyte’s broad line of enriched/purified collagenase and purified neutral protease products are rigorously characterized to ensure lot to lot consistency. Collagenase Gold is a low cost product that performs similarly to the higher priced

Evolution of Enzyme Requirements for Human Islet Isolation, Link to a Recent Review

The quality of collagenase used for mammalian islet isolation has been a focus of clinical research scientists soon after Ballinger and Lacy demonstrated that islet transplantation could correct chemically induced diabetes in inbred strains of rats (1). Clostridium histolyticum collagenase became a critical reagent because of the need to isolate sufficient numbers of functional islets

Truth vs. Fiction: Thermolysin & Human Islet Isolation

More human islet isolations have been performed using purified collagenase and thermolysin than any other purified enzyme mixture (1). Several reports have shown the adverse effects when using thermolysin for human islet isolation (2, 3). This blog post reviews the truth and fiction when using thermolysin for human islet isolation. It will also briefly discuss

New Insights for Using Purified Enzymes in Human Islet Isolation

The introduction of Liberase™ HI Purified Enzyme Blend in 1994 to improve the consistency of enzymes used to recover human pancreatic islets for subsequent transplantation contributed to the growth of clinical islet transplantation.(1)  Liberase™ HI set a benchmark for comparing the performance of all subsequent products used for this application.(2, 3) A series of reports

Variability in Islet Yields May be Due to the Molecular Form of Class 1 Collagenase

The transition of enzyme suppliers in 2007 from Liberase™ HI to Nordmark/Serva’s NB-1 collagenase & NB protease for isolation of human islets used in Collaborative Islet Transplantation consortium clinical trial was problematic for some expert islet labs but not for others. During this period, the number of allo-islet transplantations performed in North America was at

Learning from Lot Qualification

Current Lot Qualification Process Lot selection of traditional Clostridium histolyticum collagenase (i.e., crude or enriched) products is a time consuming process. Typically, you request small samples from one or more new lots of the collagenase product you currently use.  You evaluate their ability to recover sufficient numbers of functional cells, as compared to your current

Simple Approach to Optimize Collagenase Enzyme Mixtures to Improve Cell Recovery

Current State of the Art No simple method exists for defining optimal collagenase enzyme formulations for the recovery of primary cells from tissue. The unknown biochemical composition of traditional crude or enriched collagenase products, combined with variability in enzyme activities between each lot of the same product, precludes their use for this application. One option

Setting a New Paradigm for Low Cost Collagenase

The Current Paradigm for Collagenase Crude collagenase is impure, as reflected by the brown to black color of the product. Biochemical heterogeneity between different lots of crude collagenase is due to impurities and is reflected by inconsistent results after use in cell isolation procedures. Collagenase products are often sold as different “Types” that reflect their

Does Collagenase Truly Deserve Top Billing as the Key Enzyme for Isolating Primary Cells?

Collagenase Quality For many years, experts in cell isolation have stated the importance of collagenase quality for successful cell isolation.1,2 What does collagenase quality mean? Many investigators have found it difficult to correlate the enzymatic analysis data summarized on a Certificate of Analysis to the performance of a specific lot of crude or enriched collagenase

3 Factors May Explain Why You Cannot Find a “Good Lot” of Collagenase

Over the past several years, you may have found it difficult to find a good lot of collagenase from suppliers. If you’ve not experienced this in the past, you may wonder why this is happening now. Based upon my experience in collagenase product manufacture, and my interest in how business practices change over time, I

Collagenase Color: What does it tell you About Product Quality?

  What causes variation in the color of traditional collagenase products? Traditional collagenase products can range in color from a light tan to dark brown. Color is directly influenced by the collagenase manufacturing process. Collagenases and neutral proteases are the key enzymes in these products principally responsible for degrading extracellular matrix and in turn, releasing