VitaCyte is an expert in highly-purified enzymes.
Our partner VitaCyte offers highly-purified tissue dissociation enzymes for basic research as well as for cell therapy. The US-based enzyme specialist presents the broadest line of high-quality purified cell and tissue dissociation enzymes e.g. collagenases, protease on the market. Some are also in GMP quality available.
Facts, References about VitaCyte.
Low Cost, Enriched Collagenase-Purified Protease Enzyme Mixtures Successfully Used for Human Islet Isolation
Loganathan G, Hughes MG, Szot GL, Smith KE, Hussain A, Collins DR, Green ML, Dwulet FE, Williams SK, Papas KK, McCarthy RC, Balamurugan AN. OBM Transplantation 2019;3(2):14; doi:10.21926/obm.transplant.1902064.
Evolution of Enzyme Requirements for Human Islet Isolation
McCarthy RC, Green ML, Dwulet FE. OBM Transplantation 2018;2(4):024; doi:10.21926/obm.transplant.1804024.
Improved Recovery of Human Islets From Young Donor Pancreases Utilizing Increased Protease Dose to Collagenase for Digesting Peri‐Islet Extracellular Matrix
Loganathan G, Subhashree V, Narayanan S, et al. Am J Transplant. 2018;00:1–13. https://doi.org/10.1111/ajt.15111.
Beneficial Effect of Recombinant rC1rC2 Collagenases on Human Islet Function: Efficacy of Low‐Dose Enzymes on Pancreas Digestion and Yield
Loganathan G, Subhashree V, Breite AG, et al. Am J Transplant. 2018;18:478–485. https://doi.org/10.1111/ajt.14542.
Identifying Effective Enzyme Activity Targets for Recombinant Class I and Class II Collagenase for Successful Human Islet Isolation
N. Balamurugan, Appakalai & Green, Mike & Breite, Andrew & Loganathan, Gopalakrishnan & Wilhelm, Josh & Tweed, Ben & Vargova, Lenka & Lockridge, Amber & Kuriti, Manikya & Hughes, Jr, Michael & Williams, Stuart & J. Hering, Bernhard & E. Dwulet, Francis & Mccarthy, Robert. (2015). Transplantation Direct. 2. 1. 10.1097/TXD.0000000000000563.
Characterization and Functional Assessment of Clostridium Histolyticum Class I (C1) Collagenases and the Synergistic Degradation of Native Collagen in Enzyme Mixtures Containing Class II (C2) Collagenase
A.G. Breite, R.C. McCarthy, F.E. Dwulet. (2011). Transplantation Proceedings. 43. 3171-5. https://doi.org/10.1016/j.transproceed.2011.09.059.
Successful Human Islet Isolation and Transplantation Indicating the Importance of Class 1 Collagenase and Collagen Degradation Activity Assay
A. N. Balamurugan, Andrew G. Breite, Takayuki Anazawa, Gopalakrishnan Loganathan, Joshua J. Wilhelm, Klearchos K. Papas, Francis E. Dwulet, Robert C. McCarthy, Bernhard J. Hering. (2010). Transplantation. 89(8): 954–961. doi: 10.1097/TP.0b013e3181d21e9a.
Development and Application of Purified Tissue Dissociation Enzyme Mixtures for Human Hepatocyte Isolation
Roberto Gramignoli, Michael L. Green, Veysel Tahan, et al. (2012): DOI:10.3727/096368911X600939
Effectiveness of Different Molecular Forms of C. histolyticum Class I Collagenase to Recover Islets
Michael L. Green, Andrew G. Breite, Caleb A. Beechler, Francis E. Dwulet & Robert C. McCarthy (2017): Islets, DOI: 10.1080/19382014.2017.1365996
A New Enzyme Mixture to Increase the Yield and Transplant Rate of Autologous and Allogeneic Human Islet Products
Balamurugan AN, Loganathan G, Bellin MD, Wilhelm JJ, Harmon J, Anazawa T, et al. Transplantation. 2012;93(7):693-702.
Tissue Dissociation Enzymes for Isolating Human Islets for Transplantation: Factors to Consider in Setting Enzyme Acceptance Criteria
McCarthy RC, Breite AG, Green ML, Dwulet FE. Transplantation. 2011;91(2):137-45.
Tissue Dissociation Enzyme Neutral Protease Assessment
Breite A, Dwulet F, McCarthy R. Transplantation proceedings. 2010;42(6):2052-4.
Development and Characterization of a Collagen Degradation Assay to Assess Purified Collagenase Used in Islet Isolation
McCarthy RC, Spurlin B, Wright MJ, Breite AG, Sturdevant LK, Dwulet CS, et al. Transplant Proc. 2008;40(2):339-42.
Optimizing Porcine Islet Isolation to Markedly Reduce Enzyme Consumption Without Sacrificing Islet Yield or Function
Holdcraft RW, Green ML, Breite AG, Circle L, Meyer ED, Adkins H, et al. Transplantation Direct. 9000;Online Now.
Mouse Islet of Langerhans Isolation Using a Combination of Purified Collagenase and Neutral Protease
Stull, Natalie & Breite, Andrew & Mccarthy, Robert & Tersey, Sarah & Mirmira, Raghavendra. (2012). Journal of visualized experiments : JoVE. 10.3791/4137.
Frequently asked questions
Do I have to do the lot qualification procedure?
Yes. Lot qualification is important. There are variations in enzymatic/biochemical composition between various batches of poorly defined, crude, or enriched collagenase products.
Substitution of poorly defined collagenase with defined collagenase typically involves a single set of batch qualification experiments. Once the enzyme formulation and dosage are defined, the same product and dosage can be used in future cell isolations with different batches of the same product. The strict control of collagenase and protease activity of these products allows the use of different lots with prequalification.
Can I use the protease I currently take for cell isolation?
Yes. The additional protease taken for cell isolation depends on the specifications of the enzyme suppliers. It may be that other proteases work as well, but you have to find out by experimentation. Three different proteases are used to isolate human islets: Thermolysin, BP protease (a dispase-equivalent enzyme), and neutral C. histolyticum protease.
However, it may not be easy to replace the protease as the kinetics of digestion can be very different for the various general proteases. If you have questions selecting a protease, please contact us, we are one call away.
What factors should I control to make sure the cells are released properly from the tissue?
The successful isolation of cells demands excessive collagen degradation activity and the selection of an adequate neutral protease at an optimal dosage. We assume:
Enriched or purified collagenase has limited specificity to degrade only native collagen or gelatin. Excess activity does not affect the degradation of collagen and does not damage cells.
Neutral protease is required for the degradation of extracellular matrix proteins, e.g. collagen or gelatin. Proteases can also harm proteins on the cell membrane.
Bottom line: Picking the right protease at the right dose is essential for successful results.
I forgot to store my enzymes in the freezer overnight. Can I still use them?
It depends: The DE Collagenase, Collagenase Gold, Collagenase Gold Plus, BP Protease, and CIzyme AS products are stable for several days at room temperatures. For other products, please contact us, we are just one call away.
