Applications

The quality of tissue dissociation enzymes used in the cell isolation procedure are a key reagent responsible for the success of most primary cell isolation procedures. This is best illustrated by the impact purified tissue dissociation enzymes had on the improvement of human islet recovery1,2 that in turn indirectly contributed to the success of islet transplantation to manage adult patients with type 1 diabetes3.

VitaCyte has applied extensive biochemical knowledge of enzymes to the manufacture and application of tissue dissociation enzymes to offer customers three new application specific, tissue dissociation enzyme mixtures designed to isolate rodent islets, human hepatocytes, or human adipose stromal cells. Each product contains an optimal amount of purified Clostridium histolyticum collagenase and a bacterial derived neutral protease designed to recover high yields of functional cells. These formulations have been validated by experts in the field with these data shared in presentations at scientific meetings or in publications4-6.

The benefits of using these defined application specific products over crude or other ill defined collagenase containing tissue dissociation enzyme products include:

  • Consistency: rigorous characterization of the purified enzymes by functional enzyme assays results in the preparation of a consistent enzyme mixture
  • Convenience: the product contains the appropriate amount of enzymes for each specific application, which is ready to use after reconstituting
  • Cost savings: lot qualification is eliminated because purification removes the problem of lot to lot variability
  • Communication: the use of purified, functionally characterized enzymes enables these data to be shared with other laboratories so that results are confidently compared
  • Control: the adoption and use of defined tissue dissociation enzyme products simplifies translation of research results into a formalized quality system mandatory for preclinical studies

We welcome your feedback on use of our products in cell isolation procedures and encourage you to provide recommendations for new products as we expand this product line in the future.

Rodent Islets Human Hepatocytes Human Adipose Derived Stromal Cells

Reference List

1.   Linetsky E., Bottino R., Lehmann R., Alejandro R., Inverardi L., and Ricordi C. (1997) Improved human islet isolation using a new enzyme blend, liberase. Diabetes 46, 1120-3.

2.   Olack B.J., Swanson C.J., Howard T.K., and Mohanakumar T. (1999) Improved method for the isolation and purification of human islets of langerhans using Liberase enzyme blend. Hum Immunol 60, 1303-9.

3.   Shapiro A.M., Lakey J.R., Ryan E.A., Korbutt G.S., Toth E., Warnock G.L., Kneteman N.M., and Rajotte R.V. (2000) Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. N Engl J Med 343, 230-8.

4.   Gramignoli R. , Green M., Tahan V., Dorko K., Skvorak K., Marongiu F., Zao, and W e.a. (2011) Development and application of purified tissue dissociation enzyme mixtures for human hepatocyte isolation. Cell Transplantation. ePub

5.   Breite A.G., Stull N., McCarthy R., Mirmira R., and Dwulet F. (2011) Purified tissue dissociation enzyme performance in isolating mouse islets. Joint Congress Cell Transplantation and International Xenotrasplantation Societies Abstract PO50.

6.   Dale J.R., Breite D., Clayton L., Dwulet F., McCarthy R., Hoying J.B., and Williams S. (2011) Impact of enzyme composition on adipose derived stromal vacular fraction cell isolation. IFATS Annual meeting, Abstract 137.