World Pharmaceutical Frontiers talk to Roche Applied Science's Dr Cordula Jany, international product manager, and Dr Peter Hloch, market segment leader pharma biotech, about how Liberase Research Grade enzymes can maximise the yield of isolated cells that are viable and functionally active.
Dissociated tissues have been used in a variety of tissue engineering research projects. Tissue dissociation methods vary as widely as the tissues of interest. Often, enzymes will digest the extracellular matrix and release individual cells for subsequent harvesting and culturing.
Cordula Jany: The goal of cell isolation is to maximise the yield of dissociated cells that are viable and functionally active. Liberase Research Grade products are blends of highly purified enzymes that improve the quality and reproducibility of tissue dissociation, producing higher cell viability and cell function. The five different blends are: Liberase DL, Liberase TL, Liberase TM, Liberase TH and Liberase DH. The D indicates the enzyme blend contains Dispase®, the T blends contain Thermolysin. L, M and H refer to low, medium and high-relative levels of Dispase or Thermolysin.
CJ: Liberase Research Grade blends combine different ratios of highly purified, non-clostridial neutral proteases - Dispase or Thermolysin, with ultra-pure collagenase I/II enzymes (>95% homogeneity) produced using proprietary manufacturing. Roche's Liberase products replace traditional collagenases, which are characterised by crude and variable fermentation by-products of the bacteria Clostridium histolyticum.
CJ: Our second-generation Liberase blends are purified to a high degree, resulting in reduced clostripain and trypsin activity. These optimised production procedures allow our customers gentler dissociation with increased cell yield, viability and function. Thereby, the outcome from a given tissue sample is maximised. Because samples often originate from patients, they are difficult to obtain and are very valuable. Using a specific Liberase blend, more functional cells are isolated from a patient sample, and time and effort are saved.
Liberase Research Grade blends show high lot-to-lot consistency. Each lot is produced using stringent quality controls for consistent performance. In addition, our customers can easily transition from research applications to the clinical implementation, starting with Roche's Liberase research-grade blends using the equivalent GMP-grade Liberase later.
Peter Hloch: Yes, Roche Applied Science Custom Biotech also offers highly purified Liberase GMP Grade enzyme blends for tissue dissociation. These products are specifically developed to maximise yield, viability and safety in a variety of applications, including human pancreas islet isolation, tissue-derived adult stem cell isolation, tissue engineering, and human and animal cell isolation and cultivation.
Liberase GMP Grade products have the same blends of Collagenase I/II and neutral protease, but are manufactured in compliance with GMP guidelines. The production process is documented according to GMP guidelines, using rules governing medicinal products in the European Union. Filing the enzymes in the US Pharmacopoeia is underway to support customers in their filing processes with authorities. GMP Liberase blends are free of mammalian or avian tissue-derived raw materials.
CJ: A detailed guide for different tissue dissociation applications is available at www.collagenase.com. Recommended working concentrations are provided, as well as how to transition from Roche's first-generation Liberase enzyme blends and traditional collagenases. There is also a Liberase Research Grade Selection Kit, with quantities of each Liberase Research Grade blend to identify the best product for specific applications.