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Using Self-Assembled Monolayers to Pattern ECM Proteins and Cells on Subtrates

  • Protocol
Extracellular Matrix Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 139))

Abstract

We present a method that uses microcontact printing of alkanethiols on gold to generate patterned substrates presenting “islands” of extracellular matrix (ECM) surrounded by nonadhesive regions such that single cells attach and spread only on the adhesive regions. We have used this micropatterning technology to demonstrate that mammalian cells can be switched between growth and apoptosis programs in the presence of saturating concentrations of growth factors by either promoting or preventing cell spreading (1). From the perspective of fundamental cell biology, these results suggested that the local differentials in growth and viability that are critical for the formation of complex tissue patterns may be generated by local changes in cell-ECM interactions. In the context of cell culture technologies, such as bioreactors and cellular engineering applications, the regulation of cell function by cell shape indicates that the adhesive microenvironment around cells can be carefully optimized by patterning a substrate in addition to using soluble factors (2). Micropatterning technology will play a central role both in our understanding how ECM and cell shape regulate cell physiology and in facilitating the development of cellular biosensor and tissue engineering applications (3-5).

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References

  1. Chen, C. S., Mrksich, M., Huang, S., Whitesides, G. M., and Ingber, D. E. (1997) Geometric control of cell life and death. Science 276, 1425–1428.

    Article  PubMed  CAS  Google Scholar 

  2. Chen, C. S., Mrksich, M., Huang, S., Whitesides, G. M., and Ingber, D. E. (1998) Micropatterned surfaces for control of cell shape, position, and function. Biotech. Prog. 14, 356–363.

    Article  CAS  Google Scholar 

  3. Singhvi, R., Kumar, A., Lopez, G., Stephanopoulos, G. N., Wang, D. I. C., Whitesides, G. M., and Ingber, D. E. (1994) Engineering cell shape and function. Science 264, 696–698.

    Article  PubMed  CAS  Google Scholar 

  4. Hammarback, J. A., Palm, S. L., Furcht, L. T., and Letourneau, P. C. (1985) Guidance of neurite outgrowth by pathways of substratum-adsorbed laminin. J. Neurosci. Res. 13, 213–220.

    Article  PubMed  CAS  Google Scholar 

  5. Healy, K. E., Lom, B., and Hockberger, P. E. (1994) Spatial distribution of mammalian cells dictated by material surface chemistry. Biotech. Bioeng. 43, 792–800.

    Article  CAS  Google Scholar 

  6. Letourneau, P. C. (1975) Cell-to-substratum adhesion and guidance of axonal elongation. Devel. Biol. 44, 92–101.

    Article  CAS  Google Scholar 

  7. O’Neill, C., Jordan, P., and Riddle, P. (1990) Evidence for two distinct mechanisms of anchorage stimulation in freshly explanted and 3T3 swiss mouse fibroblasts. J. Cell Sci. 95, 577–586.

    PubMed  Google Scholar 

  8. Bhatia, S. N., Toner, M., Tompkins, R. G., and Yarmush, M. L. (1994) Selective adhesion of hepatocytes on patterned surfaces. Ann. NYAcad. Sci. 745, 187–209.

    Article  CAS  Google Scholar 

  9. Matsuda, T. and Sugawara, T. (1995) Development of surface photochemical modification method for micropatterning of cultured cells. J. Biomed. Mat. Res. 29, 749–756.

    Article  CAS  Google Scholar 

  10. Stenger, D. A., Georger, J. H., Dulcey, C. S., Hickman, J. J., Rudolph, A. S., et al. (1992) Coplanar molecular assemblies of amino-and perfluorinated alkylsilanes: characterization and geometric definition of mammalian cell adhesion and growth. J. Am. Chem. Soc. 114, 8435–8442.

    Article  CAS  Google Scholar 

  11. Britland, S., Clark, P., Connolly, P., and Moores, G. (1992) Micropatterned substratum adhesiveness: a model for morphogenetic cues controlling cell behavior. Exp. Cell Res. 198 124–129.

    Article  PubMed  CAS  Google Scholar 

  12. Kleinfeld, D., Kahler, K. H., and Hockberger, P, E. (1988) Controlled outgrowth of dissociated neurons on patterned substrates. J. Neurosci. 8, 4098–4120.

    PubMed  CAS  Google Scholar 

  13. Whitesides, G. M. and Gorman, C. B. (1995) Self-assembled monolayers: Models for organic surface chemistry, in Handbook of Surface Imaging and Visualization (Hubbard, A. T., ed.), CRC, Boca Raton, FL, pp. 713–732.

    Google Scholar 

  14. Tidwell, C. D., Ertel, S. I., and Ratner, B. D. (1997) Endothelial cell growth and protein adsorption on terminally functionalized, self-assembled monolayers of alkanethiolates on gold. Langmuir 13, 3404–3413.

    Article  CAS  Google Scholar 

  15. Roberts, C., Chen, C., Mrksich, M., Martichonok, V., Ingber, D., and Whitesides, G. M. (1998) Using mixed self-assembled monolayers presenting RGD and (EG)3OH groups to characterize long-term attachment of bovine capillary endothelial cells to surfaces. J. Am. Chem. Soc. 120, 6548–6555.

    Article  CAS  Google Scholar 

  16. Rao, J. and Whitesides, G. M. (1997) Tight binding of a dimeric derivative of vancomycin with dimeric l-Lys-d-Ala-d-Ala. J. Am. Chem. Soc. 119, 10,286–10,290.

    Article  CAS  Google Scholar 

  17. Wilbur, J. L., Kim, E., Xia, Y., and Whitesides, G. (1995) Lithographic molding: a convenient route to structures with sub-micrometer dimensions. Adv. Mat. 7, 649–652.

    Article  CAS  Google Scholar 

  18. Mrksich, M., Dike, L. E., Tien, J., Ingber, D. E., and Whitesides, G. M. (1997) Using microcontact printing to pattern the attachment of mammalian cells to self-assembled monolayers of alkanethiolates on transparent films of gold and silver. Exp. Cell Res. 235, 305–313.

    Article  PubMed  CAS  Google Scholar 

  19. López, G. P., Biebuyck, H. A., Härter, R., Kumar, A., and Whitesides, G. M. (1993) Fabrication and imaging of two-dimensional patterns of proteins adsorbed on self-assembled monolayers by scanning electron micorscopy. J. Am. Chem. Soc. 115, 10,774–10,781.

    Article  Google Scholar 

  20. López, G. P., Albers, M. W., Schreiber, S. L., Carroll, R. W., Peralta, E., and Whitesides, G. M. (1993) Convenient methods for patterning the adhesion of mammalian cells to surfaces using self-assembled monolayers of alkanethiolates on gold. J. Am. Chem. Soc. 115, 5877–5878.

    Article  Google Scholar 

  21. Mrksich, M., Chen, C. S., Xia, Y., Dike, L. E., Ingber, D. E., and Whitesides, G. M. (1996) Controlling cell attachment on contoured surfaces with self-assembled monolayers of alkanethiolates on gold. Proc. Natl. Acad. USA 93, 10,775–10,778.

    Article  CAS  Google Scholar 

  22. DiMilla, P., Folkers, J. P., Biebuyck, H. A., Harter, R., Lopez, G., and Whitesides, G. M. (1994) Wetting and protein adsorption of self-assembled monolayers of alka nethiolates supported on transparent films of gold. J. Am. Chem. Soc. 116, 2225–2226.

    Article  CAS  Google Scholar 

  23. Xia, Y., Mrksich, M., Kim E., and Whitesides, G. M. (1995) Microcontact printing of siloxane monolayers on the surface of silicon dioxide, and its appliction in microfabrication. J. Am. Chem. Soc. 117, 9576–9577.

    Article  CAS  Google Scholar 

  24. Madou, M. (1997) In, Fundamentals of Microfabrication, 1st ed. CRC, New York, pp. 1–52.

    Google Scholar 

  25. Zubrick, J. W. (1992) The Organic Chem Lab Survival Manual, 3rd ed. Wiley, New York.

    Google Scholar 

  26. Pale-Grosdemange, C., Simon, E. S., Prime, K. L., and Whitesides, G. M. (1991) Formation of self-assembled monolayers by chemisorption of derivatives of oligo(ethylene glycol) of structure HS(CH2)11(OCH2CH2)mOH on gold. J. Am. Chem. Soc. 113, 12–20.

    Article  CAS  Google Scholar 

  27. Prime, K. L. and Whitesides, G. M. (1991) Self-assembled organic monolayers: model systems for studying adsorption of proteins at surfaces. Science 252, 1164–1167.

    Article  CAS  Google Scholar 

  28. Prime, K. L. and Whitesides, G. M. (1993) Adsorption of proteins onto surfaces containing end-attached oligo(ethylene oxide): a model system using selfassembled monolayers. J. Am. Chem. Soc. 115, 10714–10721.

    Article  CAS  Google Scholar 

  29. Zhao, X. M., Wilbur, J. L., and Whitesides, G. M. (1996) Using two-stage chemical amplication to determine the density of defects in self-assembled monolayers of alkanethiolates on gold. Langmuir 12, 3257–3264.

    Article  CAS  Google Scholar 

  30. Bain, C. D., Biebuyck, H. A., and Whitesides, G. M. (1989) Comparison of selfassembled monolayers on gold: coadsorption of thiols and disulfides. Langmuir 5, 723–727.

    Article  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Childrens Hospital, Harvard Medical School, Boston, MA

    Christopher S. Chen, Emanuele Ostuni, George M. Whitesides & Donald E. Ingber

Authors
  1. Christopher S. Chen
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  2. Emanuele Ostuni
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  3. George M. Whitesides
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  4. Donald E. Ingber
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Editor information

Editors and Affiliations

  1. The Wellcome Trust Centre for Cell-Matrix Research, School of Biological Sciences, University of Manchester, Manchester, UK

    Charles H. Streuli  & Michael E. Grant  & 

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© 2000 Humana Press Inc.

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Chen, C.S., Ostuni, E., Whitesides, G.M., Ingber, D.E. (2000). Using Self-Assembled Monolayers to Pattern ECM Proteins and Cells on Subtrates. In: Streuli, C.H., Grant, M.E. (eds) Extracellular Matrix Protocols. Methods in Molecular Biology™, vol 139. Humana Press. https://doi.org/10.1385/1-59259-063-2:209

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  • DOI: https://doi.org/10.1385/1-59259-063-2:209

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-624-6

  • Online ISBN: 978-1-59259-063-6

  • eBook Packages: Springer Protocols

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