Advertisement

Cellular and Molecular Bioengineering

, Volume 7, Issue 1, pp 122–135 | Cite as

The Number of Lines a Cell Contacts and Cell Contractility Drive the Efficiency of Contact Guidance

  • Nicholas R. Romsey
  • Yue Hou
  • Laura Lara Rodriguez
  • Ian C. Schneider
Article

Abstract

Cell migration is an important biological function that impacts many physiological and pathological processes. Often migration is directed along aligned fibers of collagen at different densities, a process called contact guidance. However, cells adhere to other components in the extracellular matrix, possibly affecting migrational behavior. Additionally, changes in intracellular contractility are well known to affect random migration, but its effect on contact guidance is less known. This study examines differences in directed migration in response to variations in the spacing of collagen, non-specific background adhesion strength and myosin II-mediated contractility. Collagen was microcontact printed onto glass substrates and timelapse live-cell microscopy was used to measure migration characteristics. Increasing the number of lines a cell contacts or decreasing contraction led to decreases in directionality, but speed changes were context dependent. This suggests that while cell migration speed is a biphasic function of contractility, directionality appears to be a monotonic, increasing function of contractility. Thus, increasing the number of lines a cell contacts or decreasing contractility degrades the contact guidance fidelity.

Keywords

Directed cell migration Carcinoma Microcontact printing Collagen Paxillin and blebbistatin 

Abbreviations

ECM

Extracellular matrix

Col

Collagen type I

PLL-PEG

Poly-l-lysine polyethylene glycol

PLL

Poly-l-lysine

FN

Fibronectin

FA

Focal adhesion

Notes

Acknowledgments

The authors thank Caroline Zahler for initiating the project during an NSF REU (EEC-1156933) experience and Dave Schmidt and Yong Luo for technical help. The authors acknowledge support from the Roy J. Carver Charitable Trust for general project funding and from NSF ARI-R2 (CMMI-0963224) for funding the renovation of the research laboratories used for these studies.

Supplementary material

12195_2013_299_MOESM1_ESM.avi (3.7 mb)
Video of the cell shown in Fig. 1A. Collagen lines are presented as green. The length of the video spans roughly 10 h. (AVI 3806 kb)
12195_2013_299_MOESM2_ESM.avi (5.3 mb)
Video of the cell shown in Fig. 2A. Collagen lines are presented as green. The length of the video spans roughly 10 h.(AVI 5386 kb)
12195_2013_299_MOESM3_ESM.avi (2.7 mb)
Video of the cell shown in Fig. 2B. Collagen lines are presented as green. The length of the video spans roughly 6 h. AVI 2801 kb)
12195_2013_299_MOESM4_ESM.avi (3.1 mb)
Video of the cell shown in Fig. 2C. Collagen lines are presented as green. The length of the video spans roughly 7 h. (AVI 3182 kb)
12195_2013_299_MOESM5_ESM.avi (11.6 mb)
Video of the cell shown in Fig. 4A. Collagen lines are presented as red and paxillin-EGFP as green. The length of the video spans roughly 1 h. (AVI 11920 kb)
12195_2013_299_MOESM6_ESM.avi (13.7 mb)
Video of the cell shown in Fig. 4B. Collagen lines are presented as red and paxillin-EGFP as green. The length of the video spans roughly 1 h. (AVI 14047 kb)
12195_2013_299_MOESM7_ESM.avi (2.7 mb)
Video of the cell shown in Fig. 8A. Collagen lines are presented as green. The length of the video spans roughly 9 h. (AVI 2742 kb)
12195_2013_299_MOESM8_ESM.avi (3.6 mb)
Video of the cell shown in Fig. 8B. Collagen lines are presented as green. The length of the video spans roughly 8 h.(AVI 3686 kb)
12195_2013_299_MOESM9_ESM.avi (1003 kb)
Video of the cell shown in Fig. 8C. Collagen lines are presented as green. The length of the video spans roughly 8 h. (AVI 1002 kb)
12195_2013_299_MOESM10_ESM.avi (3.4 mb)
Video of the cell shown in Fig. 8D. Collagen lines are presented as green. The length of the video spans roughly 6 h. (AVI 3441 kb)
12195_2013_299_MOESM11_ESM.avi (10.3 mb)
Video of a cell migrating on 3x5 collagen lines backfilled with fibronectin from Fig. S2. Fibronectin backfill is presented as green. The length of the video spans roughly 7 h. (AVI 10563 kb)
12195_2013_299_MOESM12_ESM.avi (9.4 mb)
Video of a cell migrating on 3x10 collagen lines backfilled with fibronectin from Fig. S2. Fibronectin backfill is presented as green. The length of the video spans roughly 7 h. (AVI 9581 kb)
12195_2013_299_MOESM13_ESM.tif (140 kb)
Fig. S1 Cell Adhesion Strength: Cells were plated on unpatterned substrates as described in the “Materials and Methods” section and spun in media. Cell number was counted before and after spinning and the fraction remaining was normalized to the largest fraction (Col:PLL) (n samples = 2 and n images = 10). Error bars represent 95% confidence intervals. (TIFF 140 kb)
12195_2013_299_MOESM14_ESM.tif (972 kb)
Fig. S2 Directionality and cell migration speed on Col:FN substrates: A. Directionality and B. cell migration speed are shown for various line spacing widths and backfill molecules. Data is similar to Fig. 5 with the addition of FN backfilled substrates. Col:FN: 3x5 μm (n cells = 14 and n substrates = 1) and 3x10 μm (n cells = 15 and n substrates = 1). C. Directionality and D. cell migration speed is plotted as a function of the number of lines over which a cell spans. Col:FN: low (n cells = 6 and n substrates = 2), medium (n cells = 8 and n substrates = 1) and high (n cells = 11 and n substrates = 2). Number of lines (< 3, low; 3-5, medium; > 5, high). Col:PLL-PEG (circles), Col:PLL (squares) and Col:FN (triangles). Lines guide the eyes and error bars are 95% confidence intervals. (TIFF 973 kb)

References

  1. 1.
    Alexandrova, A. Y., K. Arnold, S. Schaub, J. M. Vasiliev, J. J. Meister, A. D. Bershadsky, and A. B. Verkhovsky. Comparative dynamics of retrograde actin flow and focal adhesions: formation of nascent adhesions triggers transition from fast to slow flow. PLoS One 3:9, 2008.CrossRefGoogle Scholar
  2. 2.
    Amyot, F., A. Small, H. Boukari, D. Sackett, J. Elliott, D. McDaniel, A. Plant, and A. Gandjbakhche. Thin films of oriented collagen fibrils for cell motility studies. J. Biomed. Mater. Res. Part B 86B:438–443, 2008.CrossRefGoogle Scholar
  3. 3.
    Anderson, A. R. A., A. M. Weaver, P. T. Cummings, and V. Quaranta. Tumor morphology and phenotypic evolution driven by selective pressure from the microenvironment. Cell 127:905–915, 2006.CrossRefGoogle Scholar
  4. 4.
    Borghi, N., M. Lowndes, V. Maruthamuthu, M. L. Gardel, and W. J. Nelson. Regulation of cell motile behavior by crosstalk between cadherin- and integrin-mediated adhesions. Proc. Natl Acad. Sci. U.S.A. 107:13324–13329, 2010.CrossRefGoogle Scholar
  5. 5.
    Branch, D. W., J. M. Corey, J. A. Weyhenmeyer, G. J. Brewer, and B. C. Wheeler. Microstamp patterns of biomolecules for high-resolution neuronal networks. Med. Biol. Eng. Comput. 36:135–141, 1998.CrossRefGoogle Scholar
  6. 6.
    Cai, Y. F., N. Biais, G. Giannone, M. Tanase, G. Y. Jiang, J. M. Hofman, C. H. Wiggins, P. Silberzan, A. Buguin, B. Ladoux, and M. P. Sheetz. Nonmuscle myosin IIA-dependent force inhibits cell spreading and drives F-actin flow. Biophys. J. 91:3907–3920, 2006.CrossRefGoogle Scholar
  7. 7.
    Clark, P., P. Connolly, and G. R. Moores. Cell guidance by micropatterned adhesiveness in vitro. J. Cell Sci. 103:287–292, 1992.Google Scholar
  8. 8.
    Csucs, G., K. Quirin, and G. Danuser. Locomotion of fish epidermal keratocytes on spatially selective adhesion patterns. Cell Motil. Cytoskeleton 64:856–867, 2007.CrossRefGoogle Scholar
  9. 9.
    Desai, R. A., M. K. Khan, S. B. Gopal, and C. S. Chen. Subcellular spatial segregation of integrin subtypes by patterned multicomponent surfaces. Integr. Biol. 3:560–567, 2011.CrossRefGoogle Scholar
  10. 10.
    Dimilla, P. A., K. Barbee, and D. A. Lauffenburger. Mathematical model for the effects of adhesion and mechanics on cell migration speed. Biophys. J. 60:15–37, 1991.CrossRefGoogle Scholar
  11. 11.
    Doyle, A. D., M. L. Kutys, M. A. Conti, K. Matsumoto, R. S. Adelstein, and K. M. Yamada. Micro-environmental control of cell migration—myosin IIA is required for efficient migration in fibrillar environments through control of cell adhesion dynamics. J. Cell Sci. 125:2244–2256, 2012.CrossRefGoogle Scholar
  12. 12.
    Doyle, A. D., F. W. Wang, K. Matsumoto, and K. M. Yamada. One-dimensional topography underlies three-dimensional fibrillar cell migration. J. Cell Biol. 184:481–490, 2009.CrossRefGoogle Scholar
  13. 13.
    Dunn, G. A., and J. P. Heath. New hypothesis of contact guidance in tissue-cells. Exp. Cell Res. 101:1–14, 1976.CrossRefGoogle Scholar
  14. 14.
    Friedl, P., and D. Gilmour. Collective cell migration in morphogenesis, regeneration and cancer. Nat. Rev. Mol. Cell Biol. 10:445–457, 2009.CrossRefGoogle Scholar
  15. 15.
    Garcia, A. J., P. Ducheyne, and D. Boettiger. Quantification of cell adhesion using a spinning disc device and application to surface-reactive materials. Biomaterials 18:1091–1098, 1997.CrossRefGoogle Scholar
  16. 16.
    Gupton, S. L., and C. M. Waterman-Storer. Spatiotemporal feedback between actomyosin and focal-adhesion systems optimizes rapid cell migration. Cell 125:1361–1374, 2006.CrossRefGoogle Scholar
  17. 17.
    Hou, Y., S. Hedberg, and I. Schneider. Differences in adhesion and protrusion properties correlate with differences in migration speed under EGF stimulation. BMC Biophys. 5:8, 2012.CrossRefGoogle Scholar
  18. 18.
    Iozzo, R. V. Matrix proteoglycans: from molecular design to cellular function. Annu. Rev. Biochem. 67:609–652, 1998.CrossRefGoogle Scholar
  19. 19.
    Jiang, F. Z., H. Horber, J. Howard, and D. J. Muller. Assembly of collagen into microribbons: effects of pH and electrolytes. J. Struct. Biol. 148:268–278, 2004.CrossRefGoogle Scholar
  20. 20.
    Kandere-Grzybowska, K., C. J. Campbell, G. Mahmud, Y. Komarova, S. Soh, and B. A. Grzybowski. Cell motility on micropatterned treadmills and tracks. Soft Matter. 3:672–679, 2007.CrossRefGoogle Scholar
  21. 21.
    Kovacs, M., J. Toth, C. Hetenyi, A. Malnasi-Csizmadia, and J. R. Sellers. Mechanism of blebbistatin inhibition of myosin II. J. Biol. Chem. 279:35557–35563, 2004.CrossRefGoogle Scholar
  22. 22.
    Kuntz, R. M., and W. M. Saltzman. Neutrophil motility in extracellular matrix gels: mesh size and adhesion affect speed of migration. Biophys. J. 72:1472–1480, 1997.CrossRefGoogle Scholar
  23. 23.
    Kushiro, K., S. Chang, and A. R. Asthagiri. Reprogramming directional cell motility by tuning micropattern features and cellular signals. Adv. Mater. 22:4516–4519, 2010.CrossRefGoogle Scholar
  24. 24.
    Lauffenburger, D. A., and J. J. Linderman. Receptors: Models for Binding, Trafficking, and Signaling. New York, NY: Oxford University Press, Inc., 1993.Google Scholar
  25. 25.
    Lehnert, D., B. Wehrle-Haller, C. David, U. Weiland, C. Ballestrem, B. A. Imhof, and M. Bastmeyer. Cell behaviour on micropatterned substrata: limits of extracellular matrix geometry for spreading and adhesion. J. Cell Sci. 117:41–52, 2004.CrossRefGoogle Scholar
  26. 26.
    Levental, K. R., H. M. Yu, L. Kass, J. N. Lakins, M. Egeblad, J. T. Erler, S. F. T. Fong, K. Csiszar, A. Giaccia, W. Weninger, M. Yamauchi, D. L. Gasser, and V. M. Weaver. Matrix crosslinking forces tumor progression by enhancing integrin signaling. Cell 139:891–906, 2009.Google Scholar
  27. 27.
    Lutolf, M. P., and J. A. Hubbell. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat. Biotechnol. 23:47–55, 2005.Google Scholar
  28. 28.
    Mager, M. D., V. LaPointe, and M. M. Stevens. Exploring and exploiting chemistry at the cell surface. Nat. Chem. 3:582–589, 2011.Google Scholar
  29. 29.
    Maiuri, P., E. Terriac, P. Paul-Gilloteaux, T. Vignaud, K. McNally, J. Onuffer, K. Thorn, P. A. Nguyen, N. Georgoulia, D. Soong, A. Jayo, N. Beil, J. Beneke, J. C. H. Lim, C. P. Y. Sim, Y. S. Chu, A. Jimenez-Dalmaroni, J. F. Joanny, J. P. Thiery, H. Erfle, M. Parsons, T. J. Mitchison, W. A. Lim, A. M. Lennon-Dumenil, M. Piel, M. Thery, and W. C. R. Participants. The first world cell race. Curr. Biol. 22:R673–R675, 2012.Google Scholar
  30. 30.
    Massia, S. P., and J. A. Hubbell. Immobilized amines and basic amino-acids as mimetic heparin-binding domains of cell-surface proteoglycan-mediated adhesion. J. Biol. Chem. 267:10133–10141, 1992.Google Scholar
  31. 31.
    Matthews, J. A., G. E. Wnek, D. G. Simpson, and G. L. Bowlin. Electrospinning of collagen nanofibers. Biomacromolecules 3:232–238, 2002.Google Scholar
  32. 32.
    Mierke, C. T., D. Rosel, B. Fabry, and J. Brabek. Contractile forces in tumor cell migration. Eur. J. Cell Biol. 87:669–676, 2008.Google Scholar
  33. 33.
    Nain, A. S., J. A. Phillippi, M. Sitti, J. MacKrell, P. G. Campbell, and C. Amon. Control of cell behavior by aligned micro/nanofibrous biomaterial scaffolds fabricated by spinneret-based tunable engineered parameters (STEP) technique. Small 4:1153–1159, 2008.Google Scholar
  34. 34.
    Nayal, A., D. J. Webb, C. M. Brown, E. M. Schaefer, M. Vicente-Manzanares, and A. R. Horwitz. Paxillin phosphorylation at ser273 localizes a GIT1-PIX-PAK complex and regulates adhesion and protrusion dynamics. J. Cell Biol. 173:587–599, 2006.Google Scholar
  35. 35.
    Oneill, C., P. Jordan, P. Riddle, and G. Ireland. Narrow linear strips of adhesive substratum are powerful inducers of both growth and total focal contact area. J. Cell Sci. 95:577–586, 1990.Google Scholar
  36. 36.
    Parsons, J. T., A. R. Horwitz, and M. A. Schwartz. Cell adhesion: integrating cytoskeletal dynamics and cellular tension. Nat. Rev. Mol. Cell Biol. 11:633–643, 2010.Google Scholar
  37. 37.
    Paszek, M. J., N. Zahir, K. R. Johnson, J. N. Lakins, G. I. Rozenberg, A. Gefen, C. A. Reinhart-King, S. S. Margulies, M. Dembo, D. Boettiger, D. A. Hammer, and V. M. Weaver. Tensional homeostasis and the malignant phenotype. Cancer Cell. 8:241–254, 2005.Google Scholar
  38. 38.
    Peyton, S. R., and A. J. Putnam. Extracellular matrix rigidity governs smooth muscle cell motility in a biphasic fashion. J. Cell. Physiol. 204:198–209, 2005.Google Scholar
  39. 39.
    Pouthas, F., P. Girard, V. Lecaudey, T. B. N. Ly, D. Gilmour, C. Boulin, R. Pepperkok, and E. G. Reynaud. In migrating cells, the Golgi complex and the position of the centrosome depend on geometrical constraints of the substratum. J. Cell Sci. 121:2406–2414, 2008.Google Scholar
  40. 40.
    Provenzano, P. P., K. W. Eliceiri, J. M. Campbell, D. R. Inman, J. G. White, and P. J. Keely. Collagen reorganization at the tumor-stromal interface facilitates local invasion. BMC Med. 4:15, 2006.Google Scholar
  41. 41.
    Provenzano, P. P., K. W. Eliceiri, L. Yan, A. Ada-Nguema, M. W. Conklin, D. R. Inman, and P. J. Keely. Nonlinear optical imaging of cellular processes in breast cancer. Microsc. Microanal. 14:532–548, 2008.Google Scholar
  42. 42.
    Provenzano, P. P., D. R. Inman, K. W. Eliceiri, S. M. Trier, and P. J. Keely. Contact guidance mediated three-dimensional cell migration is regulated by Rho/ROCK-dependent matrix reorganization. Biophys. J. 95:5374–5384, 2008.Google Scholar
  43. 43.
    Rossier, O. M., N. Gauthier, N. Biais, W. Vonnegut, M. A. Fardin, P. Avigan, E. R. Heller, A. Mathur, S. Ghassemi, M. S. Koeckert, J. C. Hone, and M. P. Sheetz. Force generated by actomyosin contraction builds bridges between adhesive contacts. EMBO J. 29:1055–1068, 2010.Google Scholar
  44. 44.
    Sabeh, F., R. Shimizu-Hirota, and S. J. Weiss. Protease-dependent versus -independent cancer cell invasion programs: three-dimensional amoeboid movement revisited. J. Cell Biol. 185:11–19, 2009.Google Scholar
  45. 45.
    Sahai, E., J. Wyckoff, U. Philippar, J. E. Segall, F. Gertler, and J. Condeelis. Simultaneous imaging of GFP, CFP and collagen in tumors in vivo using multiphoton microscopy. BMC Biotechnol. 5:9, 2005.Google Scholar
  46. 46.
    Schneider, I. C., C. K. Hays, and C. M. Waterman. Epidermal growth factor-induced contraction regulates Paxillin phosphorylation to temporally separate traction generation from De-adhesion. Mol. Biol. Cell. 20:3155–3167, 2009.Google Scholar
  47. 47.
    Totsukawa, G., Y. Yamakita, S. Yamashiro, D. J. Hartshorne, Y. Sasaki, and F. Matsumura. Distinct roles of ROCK (Rho-kinase) and MLCK in spatial regulation of MLC phosphorylation for assembly of stress fibers and focal adhesions in 3T3 fibroblasts. J. Cell Biol. 150:797–806, 2000.Google Scholar
  48. 48.
    Wirtz, D., K. Konstantopoulos, and P. C. Searson. The physics of cancer: the role of physical interactions and mechanical forces in metastasis. Nat. Rev. Cancer 11:512–522, 2011.Google Scholar
  49. 49.
    Wolf, K., I. Mazo, H. Leung, K. Engelke, U. H. von Andrian, E. I. Deryugina, A. Y. Strongin, E. B. Brocker, and P. Friedl. Compensation mechanism in tumor cell migration: mesenchymal-amoeboid transition after blocking of pericellular proteolysis. J. Cell Biol. 160:267–277, 2003.Google Scholar
  50. 50.
    Worthylake, R. A., S. Lemoine, J. M. Watson, and K. Burridge. RhoA is required for monocyte tail retraction during transendothelial migration. J. Cell Biol. 154:147–160, 2001.Google Scholar
  51. 51.
    Wűrflinger, T., I. Gamper, T. Aach, and A. S. Sechi. Automated segmentation and tracking for large-scale analysis of focal adhesion dynamics. J. Microsc. 241:37–53, 2011.Google Scholar
  52. 52.
    Zaidel-Bar, R., C. Ballestrem, Z. Kam, and B. Geiger. Early molecular events in the assembly of matrix adhesions at the leading edge of migrating cells. J. Cell Sci. 116:4605–4613, 2003.Google Scholar
  53. 53.
    Zaman, M. H., L. M. Trapani, A. Siemeski, D. MacKellar, H. Y. Gong, R. D. Kamm, A. Wells, D. A. Lauffenburger, and P. Matsudaira. Migration of tumor cells in 3D matrices is governed by matrix stiffness along with cell-matrix adhesion and proteolysis. Proc. Natl Acad. Sci. U.S.A. 103:10889–10894, 2006.Google Scholar

Copyright information

© Biomedical Engineering Society 2013

Authors and Affiliations

  • Nicholas R. Romsey
    • 1
  • Yue Hou
    • 1
  • Laura Lara Rodriguez
    • 1
  • Ian C. Schneider
    • 1
    • 2
  1. 1.Department of Chemical and Biological EngineeringIowa State UniversityAmesUSA
  2. 2.Department of Genetics, Development and Cell BiologyIowa State UniversityAmesUSA

Personalised recommendations