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Culturing Cells on Flexible Substrates of High Refractive Indexes

  • You-Ren Liu (a1) and Po-Ling Kuo (a1)

Abstract

Mechanical cues in cellular microenvironment are central in directing a class of cellular behaviors such as the dynamic of cell adhesion, migration, and differentiation. Several advanced optical techniques, such as structured-illumination nano-profilometry (SINAP), have been developed for a better resolution of these dynamic processes. These techniques however require culturing cells on materials of refractive index close to that of glass, while most studies regarding the effects of mechanical cues on cellular dynamics were conducted on hydrogel-based substrates. Here we report the development of culturing substrates of tunable rigidity and refractive index suitable for SINAP studies. Polyvinyl chloride (PVC)-based substrates were mixed with a softener called Di(isononyl) Cyclohexane-1,2-Dicarboxylate (DINCH) and cured by heating. The volume ratios of PVC to DINCH were varied from 1:1 to 3:1. The Young’s modulus of the resulting substrates ranged from 18 kPa to 40 kPa. The yielded refractive indices of the composite substrates as measured by phase contrast tomography ranged from 1.47 to 1.53. Human lung adenocarcinoma cells CL1-5 were cultured on the composite substrates and cell viability was examined using the MTT assay. The dynamics of cell adhesion and filopodia activities were examined using SINAP. Preliminary results suggest that PVC based culturing substrates have a great potential in the application of SINAP based studies.

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1. Ridley, A.J., et al. ., Cell migration: integrating signals from front to back . Science , 2003. 302(5651): p. 17049.10.1126/science.1092053
2. Friedl, P. and Wolf, K., Plasticity of cell migration: a multiscale tuning model . J Cell Biol , 2010. 188(1): p. 119.10.1083/jcb.200909003
3. Guo, W.H., et al. ., Substrate rigidity regulates the formation and maintenance of tissues . Biophysical Journal , 2006. 90(6): p. 22132220.10.1529/biophysj.105.070144
4. Discher, D.E., Janmey, P., and Wang, Y.L., Tissue cells feel and respond to the stiffness of their substrate . Science , 2005. 310(5751): p. 11391143.
5. Guo, W. and Wang, Y., The substrate rigidity regulates the formation and maintenance of tissues . Molecular Biology of the Cell , 2004. 15: p. 3A3A.
6. Wang, Y.L., Traction forces and rigidity sensing of adherent cells . Conf Proc IEEE Eng Med Biol Soc , 2009. 2009: p. 333940.
7. Buxboim, A., Ivanovska, I.L., and Discher, D.E., Matrix elasticity, cytoskeletal forces and physics of the nucleus: how deeply do cells ’feel’ outside and in? J Cell Sci , 2010. 123(Pt 3): p. 297308.
8. Tzvetkova-Chevolleau, T., et al. ., The motility of normal and cancer cells in response to the combined influence of the substrate rigidity and anisotropic microstructure . Biomaterials , 2008. 29(10): p. 15411551.
9. Chan, C.E. and Odde, D.J., Traction Dynamics of Filopodia on Compliant Substrates . Science , 2008. 322(5908): p. 16871691.
10. Wang, C.C., et al. ., Asymmetric cancer-cell filopodium growth induced by electric-fields in a microfluidic culture chip . Lab Chip , 2011. 11(4): p. 6959.
11. Lee, C.H., Mong, H.Y., and Lin, W.C., Noninterferometric wide-field optical profilometry with nanometer depth resolution . Optics Letters , 2002. 27(20): p. 17731775.
12. Lin, J.Y., et al. ., Wide-field super-resolution optical sectioning microscopy using a single spatial light modulator . Journal of Optics a-Pure and Applied Optics , 2009. 11(1): p. -.
13. Cretu, A., Castagnino, P., and Assoian, R., Studying the effects of matrix stiffness on cellular function using acrylamide-based hydrogels . J Vis Exp , 2010(42).
14. Tse, J.R. and Engler, A.J., Preparation of hydrogel substrates with tunable mechanical properties . Curr Protoc Cell Biol , 2010. Chapter 10: p. Unit 10 16.10.1002/0471143030.cb1016s47
15. Kandow, C.E., et al. ., Polyacrylamide hydrogels for cell mechanics: steps toward optimization and alternative uses . Methods Cell Biol , 2007. 83: p. 2946.10.1016/S0091-679X(07)83002-0
16. Storck, J., AbDelRazek, H., and Zimmermann, E.R., Effect of polyvinyl chloride plastic on the growth and physiology of human umbilical vein endothelial cells . Biomaterials , 1996. 17(18): p. 17911794.
17. Buckley, M.R., et al. ., Mapping the depth dependence of shear properties in articular cartilage . Journal of Biomechanics , 2008. 41(11): p. 24302437.10.1016/j.jbiomech.2008.05.021
18. Choi, W., et al. ., Tomographic phase microscopy . Nature Methods , 2007. 4(9): p. 717719.
19. Wong, J.Y., et al. ., Directed movement of vascular smooth muscle cells on gradient-compliant hydrogels . Langmuir , 2003. 19(5): p. 19081913.10.1021/la026403p
20. Engler, A.J., et al. ., Matrix elasticity directs stem cell lineage specification . Cell , 2006. 126(4): p. 677689.

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