1. A Biaxial Cell Stretcher Client: Frank Yin, MD. Ph.D Group 30 Joshua Leibowitz Krista Vedvik Christopher Zarins

2. Background Cells in the body experience mechanical forces Heart Lungs Blood vessels Laboratory cell cultures should recreate physiological conditions

3. Need for a Cell Stretcher Studying the effects of mechanical force on aortic endothelial cells Orientation and organization of cells depends on stretching Controlling deformation in both directions gives the most accurate results

4. Other Uses of Stretching Cardiac—Induce alignment of stem cell derived cardiomyocytes (Tulloch, 2011) Vascular—Study formation of aneurysms (Ventikos, 2010) Respiratory—Study the effects of protein secretion (Arnold, 2006)

5. Need for Biaxial Tests Most current cell stretching experiments are uniaxial Limits the validity of results

6. Scope Design a device capable of applying 40% strain in two directions Withstands incubator conditions (37˚C, 100% humidity) Maintains calibration over a period of weeks Easy access to medium to allow for additions and changes

7. Current approaches Culture on an elastic membrane (Patten, 2005) Culture imbedded in a pliable gel (Zile, 2006) Cell culture Elastic membrane Adhesion Protein

8. FlexCell system Uses pressure to deform a circular membrane Applies radial strain

9. Calculations Taking E = 10 kPa, A 0 = 1 cm 2, L 0 = 10 cm, ΔL = 4 cm, F = 4 N Required deflection force: Motor accuracy: Assuming 1.8˚ stepper motor resolution, drive mechanism must have at least 0.2 mm deflection per degree

10. Design Schedule Task/Milestone3-Sep10-Sep17-Sep24-Sep1-Oct8-Oct15-Oct22-Oct29-Oct5-Nov12-Nov19-Nov26-Nov3-Dec10-Dec Project Initiation Project Selection *Project Scope Develop Project Plan Develop Requirements *Preliminary Oral Report *Preliminary Written Report *Web Page Initial Risk Analysis Design & Design Analysis Feasibility Report *Progress Oral Reports *Progress Written Reports *Final Oral Report *Final Written Report Project Poster Judging

11. Member Responsibilities ChrisJoshKrista Conceptualization xxx Device Components Substrate x Drive Mechanism x Controller Interface x Incubator Compatibility x Imaging Compatibility x Calibration x Risk Analysis xxx Research Feasibility xxx Literature Searches x Mathematical Parameters x Prices/Quotes x Reports Initializing x Scheduling/Labor Division x Figures x Copy-editing x Presentations Preliminary x Progress x Final x Client Interactions x x Intellectual Property x Website x

12. References Arnold, S. P. et. al. Design of a New Stretching Apparatus and the Effects of Cyclic Strain and Substratum on Mouse Lung Epithelial-12 Cells. Ann Biomed Eng, 2007 35, 1156-1164 Paten, J.A. et. al. Design and Performance of an Optically Accessible, Low-Volume, Mechanobioreactor for Long-Term Study of Living Constructs. Tis Eng, 2011 17 Part C Tulloch, NL. et al. Growth of engineered human myocardium with mechanical loading and vascular coculture. Circ Res. 2011 Jun 4;109(1):47-59 Ventikos et al. Modelling evolution and the evolving mechanical environment of saccular cerebral aneurysms. Biomech Model Mechanobiol 2011 10:109-132 Zile MR. et al. Gel stretch method: A new method to measure constitutive properties of cardiac muscle cells. Am J Physiol. 1998 Jun;274(6 Pt 2):h2188-202

13. Questions?