Hostname: page-component-8448b6f56d-dnltx Total loading time: 0 Render date: 2024-04-19T01:55:34.140Z Has data issue: false hasContentIssue false

Culturing Cells on Flexible Substrates of High Refractive Indexes

Published online by Cambridge University Press:  10 May 2012

You-Ren Liu
Affiliation:
Department of Electric Engineering, Graduate Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, Taipei, 10617, Taiwan
Po-Ling Kuo
Affiliation:
Department of Electric Engineering, Graduate Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, Taipei, 10617, Taiwan
Get access

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.

Type
Research Article
Copyright
Copyright © Materials Research Society 2012

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

1. Ridley, A.J., et al. ., Cell migration: integrating signals from front to back . Science , 2003. 302(5651): p. 17049.10.1126/science.1092053CrossRefGoogle ScholarPubMed
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.200909003CrossRefGoogle ScholarPubMed
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.105.070144CrossRefGoogle ScholarPubMed
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.CrossRefGoogle Scholar
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.Google Scholar
6. Wang, Y.L., Traction forces and rigidity sensing of adherent cells . Conf Proc IEEE Eng Med Biol Soc , 2009. 2009: p. 333940.Google ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle Scholar
9. Chan, C.E. and Odde, D.J., Traction Dynamics of Filopodia on Compliant Substrates . Science , 2008. 322(5908): p. 16871691.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle Scholar
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.CrossRefGoogle ScholarPubMed
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. -.Google Scholar
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).CrossRefGoogle ScholarPubMed
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.cb1016s47CrossRefGoogle ScholarPubMed
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)83002-0CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
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.2008.05.021CrossRefGoogle ScholarPubMed
18. Choi, W., et al. ., Tomographic phase microscopy . Nature Methods , 2007. 4(9): p. 717719.CrossRefGoogle ScholarPubMed
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/la026403pCrossRefGoogle Scholar
20. Engler, A.J., et al. ., Matrix elasticity directs stem cell lineage specification . Cell , 2006. 126(4): p. 677689.CrossRefGoogle ScholarPubMed