Abstract
Traditional methods to study normal and pathological development of tissues have been limited by difficulties in controlling experimental conditions and quantifying biological processes of interest. Here we describe methods to create microarrays of engineered tissues that enable controlled and quantitative investigations. Using soft lithography-based techniques, extracellular matrix proteins can be microcontact printed or micromolded to make two- and three-dimensional micropatterned scaffolds. The ultimate form and resulting properties of the tissue construct are dictated by the geometry of the patterned extracellular matrix components. This chapter describes elastomeric stamp fabrication, microcontact printing and micromolding of extracellular matrix proteins, cell culture in micropatterned substrata, and quantitative immunofluorescence analysis of micropatterned tissues.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Langer, R., and Vacanti, J. P. (1993) Tissue engineering. Science 260, 920–926.
O’Neill, C., Jordan, P., and Ireland, G. (1986) Evidence for two distinct mechanisms of anchorage stimulation in freshly explanted and 3t3 swiss mouse fibroblasts. Cell 44, 489–496.
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.
Whitesides, G. M., Ostuni, E., Takayama, S., Jiang, X., and Ingber, D. E. (2001) Soft lithography in biology and biochemistry. Annu. Rev. Biomed. Eng. 3, 335–373.
Nelson, C. M., Jean, R. P., Tan, J. L., Liu, W. F., Sniadecki, N. J., Spector, A. A., and Chen, C. S. (2005) Emergent patterns of growth controlled by multicellular form and mechanics. Proc. Natl. Acad. Sci. USA 102, 11594–11599.
Tien, J., Nelson, C. M., and Chen, C. S. (2002) Fabrication of aligned microstructures with a single elastomeric stamp. Proc. Natl. Acad. Sci. USA 99, 1758–1762.
Jeon, N. L., Dertinger, S. K. W., Chiu, D. T., Choi, I. S., Stroock, A. D., and Whitesides, G. M. (2000) Generation of solution and surface gradients using microfluidic systems. Langmuir 16, 8311–8316.
Ostuni, E., Kane, R., Chen, C. S., Ingber, D. E., and Whitesides, G. M. (2000) Patterning mammalian cells using elastomeric membranes. Langmuir 16, 7811–7819.
Flaim, C. J., Chien, S., and Bhatia, S. N. (2005) An extracellular matrix microarray for probing cellular differentiation. Nat. Methods 2, 119–125.
Wilson, D. L., Martin, R., Hong, S., Cronin-Golomb, M., Mirkin, C. A., and Kaplan, D. L. (2001) Surface organization and nanopatterning of collagen by dip-pen nanolithography. Proc. Natl. Acad. Sci. USA 98, 13660–13664.
Jackson, B. L., and Groves, J. T. (2007) Hybrid protein-lipid patterns from aluminum templates. Langmuir 23, 2052–2057.
Nguyen, K. T., and West, J. L. (2002) Photopolymerizable hydrogels for tissue engineering applications. Biomaterials 23, 4307–4314.
Tsang, V. L., Chen, A. A., Cho, L. M., Jadin, K. D., Sah, R. L., DeLong, S., West, J. L., and Bhatia, S. N. (2007) Fabrication of 3d hepatic tissues by additive photopatterning of cellular hydrogels. Faseb J. 21, 790–801.
Tang, M. D., Golden, A. P., and Tien, J. (2003) Molding of three-dimensional microstructures of gels. J. Am. Chem. Soc. 125, 12988–12989.
Tang, M. D., Golden, A. P., and Tien, J. (2004) Fabrication of collagen gels that contain patterned, micrometer-scale cavities. Adv. Mater. 16, 1345–1348.
Nelson, C. M., Vanduijn, M. M., Inman, J. L., Fletcher, D. A., and Bissell, M. J. (2006) Tissue geometry determines sites of mammary branching morphogenesis in organotypic cultures. Science 314, 298–300.
Fukuda, J., Khademhosseini, A., Yeo, Y., Yang, X. Y., Yeh, J., Eng, G., Blumling, J., Wang, C. F., Kohane, D. S., and Langer, R. (2006) Micromolding of photocrosslinkable chitosan hydrogel for spheroid microarray and co-cultures. Biomaterials 27, 5259–5267.
Nelson, C. M., Inman, J. L., and Bissell, M. J. (2008) Three-dimensional lithographically defined organotypic tissue arrays for quantitative analysis of morphogenesis and neoplastic progression. Nat. Protoc. 3, 674–678.
Jongpaiboonkit, L., King, W. J., Lyons, G. E., Paguirigan, A. L., Warrick, J. W., Beebe, D. J., and Murphy, W. L. (2008) An adaptable hydrogel array format for 3-dimensional cell culture and analysis. Biomaterials 29, 3346–3356.
Fukuda, J., Khademhosseini, A., Yeh, J., Eng, G., Cheng, J. J., Farokhzad, O. C., and Langer, R. (2006) Micropatterned cell co-cultures using layer-by-layer deposition of extracellular matrix components. Biomaterials 27, 1479–1486.
Bettinger, C. J., Weinberg, E. J., Kulig, K. M., Vacanti, J. P., Wang, Y. D., Borenstein, J. T., and Langer, R. (2006) Three-dimensional microfluidic tissue-engineering scaffolds using a flexible biodegradable polymer. Adv. Mater. 18, 165–169.
Golden, A. P., and Tien, J. (2007) Fabrication of microfluidic hydrogels using molded gelatin as a sacrificial element. Lab Chip 7, 720–725.
Tan, J. L., Liu, W., Nelson, C. M., Raghavan, S., and Chen, C. S. (2004) Simple approach to micropattern cells on common culture substrates by tuning substrate wettability. Tissue Eng. 10, 865–872.
Acknowledgments
This work was supported by grants from the NIH (CA128660 and GM083997), Susan G. Komen for the Cure (FAS0703855); and the David & Lucile Packard Foundation. C.M.N. holds a Career Award at the Scientific Interface from the Burroughs Wellcome Fund. E.W.G. was supported by post doctoral fellowships from the New Jersey Commission on Cancer Research and Susan G. Komen for the cure.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Gomez, E.W., Nelson, C.M. (2011). Lithographically Defined Two- and Three-Dimensional Tissue Microarrays. In: Khademhosseini, A., Suh, KY., Zourob, M. (eds) Biological Microarrays. Methods in Molecular Biology, vol 671. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-551-0_5
Download citation
DOI: https://doi.org/10.1007/978-1-59745-551-0_5
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-934115-95-4
Online ISBN: 978-1-59745-551-0
eBook Packages: Springer Protocols