Abstract
Several biological processes, including cell migration, tissue morphogenesis, and cancer metastasis, are fundamentally physical in nature; each implicitly involves deformations driven by mechanical forces. Traction force microscopy (TFM) was initially developed to quantify the forces exerted by individual isolated cells in two-dimensional (2D) culture. Here, we extend this technique to estimate the traction forces generated by engineered three-dimensional (3D) epithelial tissues embedded within a surrounding extracellular matrix (ECM). This technique provides insight into the physical mechanisms that underlie tissue morphogenesis in 3D.
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Abbreviations
- 2D:
-
Two-dimensional
- 3D:
-
Three-dimensional
- BSA:
-
Bovine serum albumin
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- DVC:
-
Digital volume correlation
- ECM:
-
Extracellular matrix
- EMT:
-
Epithelial-mesenchymal transition
- FBS:
-
Fetal bovine serum
- HBSS:
-
Hanks’ balanced salt solution
- PBS:
-
Phosphate-buffered saline
- PDMS:
-
Polydimethylsiloxane
- TFM:
-
Traction force microscopy
References
Nelson CM, Jean RP, Tan JL, Liu WF, Sniadecki NJ, Spector AA, Chen CS (2005) Emergent patterns of growth controlled by multicellular form and mechanics. Proc Natl Acad Sci U S A 102(33):11594–11599
McBeath R, Pirone DM, Nelson CM, Bhadriraju K, Chen CS (2004) Cell shape, cytoskeletal tension, and rhoa regulate stem cell lineage commitment. Dev Cell 6(4):483–495
Engler AJ, Sen S, Sweeney HL, Discher DE (2006) Matrix elasticity directs stem cell lineage specification. Cell 126(4):677–689
Lui C, Lee K, Nelson CM (2012) Matrix compliance and rhoa direct the differentiation of mammary progenitor cells. Biomech Model Mechanobiol 11(8):1241–1249
Gomez EW, Chen QK, Gjorevski N, Nelson CM (2010) Tissue geometry patterns epithelial-mesenchymal transition via intercellular mechanotransduction. J Cell Biochem 110(1):44–51
Lee K, Chen QK, Lui C, Cichon MA, Radisky DC, Nelson CM (2012) Matrix compliance regulates rac1b localization, nadph oxidase assembly, and epithelial-mesenchymal transition. Mol Biol Cell 23(20):4097–4108
Harris AK, Wild P, Stopak D (1980) Silicone rubber substrata: a new wrinkle in the study of cell locomotion. Science 208(4440):177–179
Lee J, Leonard M, Oliver T, Ishihara A, Jacobson K (1994) Traction forces generated by locomoting keratocytes. J Cell Biol 127(6 Pt 2):1957–1964
Dembo M, Oliver T, Ishihara A, Jacobson K (1996) Imaging the traction stresses exerted by locomoting cells with the elastic substratum method. Biophys J 70(4):2008–2022
Dembo M, Wang YL (1999) Stresses at the cell-to-substrate interface during locomotion of fibroblasts. Biophys J 76(4):2307–2316
Maskarinec SA, Franck C, Tirrell DA, Ravichandran G (2009) Quantifying cellular traction forces in three dimensions. Proc Natl Acad Sci U S A 106(52):22108–22113
Legant WR, Miller JS, Blakely BL, Cohen DM, Genin GM, Chen CS (2010) Measurement of mechanical tractions exerted by cells in three-dimensional matrices. Nat Methods 7(12):969–971
Nelson CM, Inman JL, Bissell MJ (2008) Three-dimensional lithographically defined organotypic tissue arrays for quantitative analysis of morphogenesis and neoplastic progression. Nat Protocols 3(4):674–678
Gjorevski N, Nelson CM (2012) Map** of mechanical strains and stresses around quiescent engineered three-dimensional epithelial tissues. Biophys J 103(1):152–162
Crocker JC, Grier DG (1996) Methods of digital video microscopy for colloidal studies. J Colloid Interface Sci 179(1):298–310
Franck C, Hong S, Maskarinec SA, Tirrell DA, Ravichandran G (2007) Three-dimensional full-field measurements of large deformations in soft materials using confocal microscopy and digital volume correlation. Exp Mech 47(3):427–438
Comsol multiphysics user's guide (2011). Comsol 4.2a. COMSOL AB, Burlington, MA.
Acknowledgements
We thank Lynn Loo for cleanroom access. This work was supported in part by grants from the NIH (GM083997 and HL110335), the David and Lucile Packard Foundation, the Alfred P. Sloan Foundation, and the Camille and Henry Dreyfus Foundation. C.M.N. holds a Career Award at the Scientific Interface from the Burroughs Wellcome Fund. N.G. was supported in part by a Wallace Memorial Honorific Fellowship.
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Piotrowski, A.S., Varner, V.D., Gjorevski, N., Nelson, C.M. (2015). Three-Dimensional Traction Force Microscopy of Engineered Epithelial Tissues. In: Nelson, C. (eds) Tissue Morphogenesis. Methods in Molecular Biology, vol 1189. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1164-6_13
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DOI: https://doi.org/10.1007/978-1-4939-1164-6_13
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1163-9
Online ISBN: 978-1-4939-1164-6
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