1. Quasi-Vivo TM Inter-connected Cell Culture Dr Kate Darley Kirkstall Ltd. Presentation April 2010

2. Quasi-Vivo TM Cell Culture Systems  Introduction to the Quasi-Vivo TM system  System data  Future directions  About Kirkstall  Access to the Quasi-Vivo TM system

3. Quasi-Vivo TM Cell Culture Systems  Launch of company, November 2006  South Yorkshire Seedcorn Fund investment July 2007  Licence with Pisa University and Collaboration begins with Sheffield University, October 2007  Proof of Concept Testing complete, June 2008  Second Round Seed Funding, September 2008  Manufacture and first validation studies complete, November 2008  First Product (kit) sales, February 2009  Launch of Contract Research Service, April 2009 About Kirkstall

4. Quasi-Vivo TM Cell Culture Systems Introduction to the Quasi-Vivo TM system

5. Quasi-Vivo TM Cell Culture Systems INTER-CONNECTED CELL CULTURE …  Cells/tissue connected in series/parallel  Nutrient/media flow-through system  Allows cell-to-cell signalling  Metabolites/drugs/compounds circulate in system  Vitality/cell differentiation maintained PROVIDES IN-VIVO LIKE (Quasi-vivo) CONDITIONS FOR CELL GROWTH

6. Quasi-Vivo TM Cell Culture Systems INTER-CONNECTED CELL CULTURE …

7. Quasi-Vivo TM Cell Culture Systems Chamber design  Minimises bubble formation and shear stress  Shape & dimensions similar to 24 well plates. Facilitates cell transfer …from seeding to the bio-module  Cell biologists familiar with conventional multi-well plates rapidly able to use...the system Proprietary design/covered by three patents

8. Quasi-Vivo TM Cell Culture Systems Experimental design  Basic - 2 chambers connected In series/parallel  Complex – 32 chambers connected in series/parallel

9. Quasi-Vivo TM Cell Culture Systems PROVIDES IN-VIVO LIKE CONDITIONS FOR CELL GROWTH? VALIDATION I  In primary hepatocytes xenobiotic gene expression is closer to in vivo.  Cells maintain differentiation over a longer periods Cell morphology staticQuasi-vivo

10. Quasi-Vivo TM Cell Culture Systems VALIDATION II - Correlation with drug metabolism/toxicity in vivo? Improved Sensitivity for Toxicity Screening: Diclofenac A non-steroidal anti- inflammatory. Dose-response curve in Human Primary hepatocytes Quasi- Vivo TM EC 50 Conventional In vitro EC 50 Clinical Trial Result

11. Quasi-Vivo TM Cell Culture Systems Access to the Quasi-Vivo TM system

12. Quasi-Vivo TM Cell Culture Systems Products: The Quasi-Vivo™ 100 Kit ACCESS TO THE TECHNOLOGY Bio-modules, reservoir bottle, connecting tubes, Peristaltic pump (Kit). Services: Contract research Flexible service, labs service available in Sheffield or in your own lab

13. Quasi-Vivo TM Cell Culture Systems WHO WE WORK WITH….  Academic groups: UK universities, France (Montpellier), Italy (Pisa), EU framework 7 – InLIVEtox  Cosmetic industry – EU ban 2009 Need for an alternative to animal testing  Chemical industry – REACH legislation Registration, Evaluation, Authorisation and restriction of CHemicals  Pharmaceutical industry- Lead discovery/lead optimization

14. KEY NOTE SPEAKERS: Prof David Tweats (University of Swansea) ‘Toxicity Testing in the 21st Century’ Prof Claus-Michael Lehr (Helmholtz-Institute, Saarland) ‘Drug delivery across biological barriers’ Dr Patrick Maurel (INSERM Montpellier) ‘Human hepatocyte culture and applications for drug discovery’ Dr David Standring (Idenix Pharma, MA, USA) ‘Preclinical drug discovery, in vitro/in vivo testing past and future’ Dr Robert Landsiedel (BASF) ‘Safety Research on Nanomaterials’ Prof Sheila Macneil (University of Sheffield) ‘3D Tissue engineering models for R&D’

15. Quasi-Vivo TM Cell Culture Systems Thanks for listening & any questions… kdarley@kirkstall.org www.kirkstall.org