Advertisement

Measurement of Intercellular Cohesion by Tissue Surface Tensiometry

  • Ramsey A. FotyEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1189)

Abstract

Intercellular adhesion plays a vital role in many biological processes including embryonic development, malignant invasion, and wound healing, and can be manipulated to generate complex structures in tissue engineering applications. Accurate measurement of the strength of intercellular adhesion is not trivial and requires methods rooted in sound physical principles. Tissue surface tensiometry (TST) rigorously quantifies intercellular cohesive energy of 3D tissue-like aggregates under physiological conditions. TST utilizes a custom-built tensiometer to compress 3D spheroids between parallel plates. The resistance to the applied force and changes in aggregate geometry are applied to the Young-Laplace equation, generating a measurement of apparent surface tension. We describe all components comprising the tensiometer and provide step by step instructions of all the key steps involved in generating spherical aggregates. We explain how tissue surface tension is calculated and provide a statistical analysis of a sample data set from 12 aggregates.

Key words

Cell adhesion Tissue liquidity Surface tension Tissue surface tensiometry Parallel plate compression 

Notes

Acknowledgements

This work was supported by NIH grant CA118755 to RAF. The author also acknowledges the seminal contribution of Malcolm S. Steinberg, Ph.D., to the development of this method.

Supplementary material

Movie 16.1

Aggregate rounding up: Here, an “irregular” fragment of embryonic chick liver was extirpated from a 3.5-day-old embryo, and placed onto the lower compression plate (LCP). The LCP had been pre-coated with poly-HEMA, preventing adhesion of the fragment to the plate. The tissue fragment was incubated overnight and filmed in real time. The movie is run at 8× speed. Note the rounding-up behavior and the significant movement of the aggregate on the plate, indicating little adhesion to the substrate (MOV 44708 kb).

Movie 16.2

Real-time chart recorder tracing as an aggregate is undergoing compression and force relaxation. The pen deflects from the zero force position at chart recorder position 10 to approximately position 52, representing an applied weight of 4.2 mg. The aggregate undergoes a fast relaxation phase in the first 45 s after compression, whereupon the force tracing begins to level off (MOV 77785 kb).

Movie 16.3

Real-time chart recorder tracing as an aggregate is decompressed after having reached force and shape equilibrium. This aggregate was under compression for approximately 2 h and 20 min. By this time, the force tracing had leveled off at the 1.9 mg position, indicating that the aggregate had reached equilibrium. Upon decompression, the chart recorder pen deflected back towards the zero force balance position. Accordingly, the F value recorded for application to the Young-Laplace equation was 1.9 mg (MOV 48674 kb).

Movie 16.4

The elastic response. An embryonic chick aggregate composed of myocardium when compressed and immediately decompressed assumes its original shape (MOV 29802 kb).

Movie 16.5

The viscous liquid response. An aggregate of embryonic chick myocardium if compressed and allowed to reach shape and force equilibrium does not assume its original spherical shape upon decompression. Note that rounding up takes place in a more gradual manner as depicted in time lapse (MOV 245564 kb).

References

  1. 1.
    Foty RA, Pfleger CM, Forgacs G, Steinberg MS (1996) Surface tensions of embryonic tissues predict their mutual envelopment behavior. Development 122(5):1611–1620PubMedGoogle Scholar
  2. 2.
    Foty RA, Steinberg MS (2005) The differential adhesion hypothesis: a direct evaluation. Dev Biol 278(1):255–263PubMedCrossRefGoogle Scholar
  3. 3.
    Schotz EM, Burdine RD, Julicher F, Steinberg MS, Heisenberg CP, Foty RA (2008) Quantitative differences in tissue surface tension influence zebrafish germ layer positioning. HFSP J 2(1):42–56PubMedCrossRefPubMedCentralGoogle Scholar
  4. 4.
    Hackett-Jones EJ, Landman KA, Newgreen DF, Zhang D (2011) On the role of differential adhesion in gangliogenesis in the enteric nervous system. J Theor Biol 287:148–159PubMedCrossRefGoogle Scholar
  5. 5.
    Kragl M, Knapp D, Nacu E, Khattak S, Maden M, Epperlein HH, Tanaka EM (2009) Cells keep a memory of their tissue origin during axolotl limb regeneration. Nature 460(7251):60–65PubMedCrossRefGoogle Scholar
  6. 6.
    Wada N (2011) Spatiotemporal changes in cell adhesiveness during vertebrate limb morphogenesis. Dev Dyn 240(5):969–978PubMedCrossRefGoogle Scholar
  7. 7.
    Foty RA, Steinberg MS (2013) Differential adhesion in model systems. WIREs Dev Biol 2:631–645CrossRefGoogle Scholar
  8. 8.
    Steinberg M (1964) The problem of adhesive selectivity in cellular interactions. In: Locke M (ed) Cellular membranes in development 22nd symposium of the society for the study of development and growth. Academic, New York, NY, pp 321–366CrossRefGoogle Scholar
  9. 9.
    Moyer WA, Steinberg MS (1976) Do rates of intercellular adhesion measure the cell affinities reflected in cell sorting and tissue spreading configurations. Dev Biol 52:246–262PubMedCrossRefGoogle Scholar
  10. 10.
    Nose A, Nagafuchi A, Takeichi M (1988) Expressed recombinant cadherins mediate cell sorting in model systems. Cell 54:993–1001PubMedCrossRefGoogle Scholar
  11. 11.
    Glasstone S, Laidler KJ, Eyring H (1941) The theory of rate processes; the kinetics of chemical reactions, viscosity, diffusion and electrochemical phenomena. McGraw-Hill Book Company, Inc., New York, NYGoogle Scholar
  12. 12.
    Duguay D, Foty RA, Steinberg MS (2003) Cadherin-mediated cell adhesion and tissue segregation: qualitative and quantitative determinants. Dev Biol 253(2):309–323PubMedCrossRefGoogle Scholar
  13. 13.
    Chu YS, Thomas WA, Eder O, Pincet F, Perez E, Thiery JP, Dufour S (2004) Force measurements in E-cadherin-mediated cell doublets reveal rapid adhesion strengthened by actin cytoskeleton remodeling through Rac and Cdc42. J Cell Biol 167(6):1183–1194PubMedCrossRefPubMedCentralGoogle Scholar
  14. 14.
    Bell GI (1978) Models for the specific adhesion of cells to cells. Science 200(4342):618–627PubMedCrossRefGoogle Scholar
  15. 15.
    Evans E, Berk D, Leung A (1991) Detachment of agglutinin-bonded red blood cells I. Forces to rupture molecular-point attachments. Biophys J 59:838–848PubMedCrossRefPubMedCentralGoogle Scholar
  16. 16.
    Steinberg MS (1996) Adhesion in development: an historical overview. Dev Biol 180(2):377–388PubMedCrossRefGoogle Scholar
  17. 17.
    Foty RA, Steinberg MS (1997) Measurement of tumor cell cohesion and suppression of invasion by E- or P-cadherin. Cancer Res 57(22):5033–5036PubMedGoogle Scholar
  18. 18.
    Jia D, Dajusta D, Foty RA (2007) Tissue surface tensions guide in vitro self-assembly of rodent pancreatic islet cells. Dev Dyn 236(8):2039–2049PubMedCrossRefGoogle Scholar
  19. 19.
    Sabari J, Lax D, Connors D, Brotman I, Mindrebo E, Butler C, Entersz I, Jia D, Foty RA (2011) Fibronectin matrix assembly suppresses dispersal of glioblastoma cells. PLoS One 6(9):e24810PubMedCrossRefPubMedCentralGoogle Scholar
  20. 20.
    Winters BS, Shepard SR, Foty RA (2005) Biophysical measurement of brain tumor cohesion. Int J Cancer 114(3):371–379PubMedCrossRefGoogle Scholar
  21. 21.
    Davies JT, Rideal EK (1963) Interfacial phenomena. Academic, New York, NYGoogle Scholar
  22. 22.
    Foty R (2011) A simple hanging drop cell culture protocol for generation of 3D spheroids. J Vis Exp (51)Google Scholar
  23. 23.
    Butler CM, Foty RA (2011) Measurement of aggregate cohesion by tissue surface tensiometry. J Vis Exp (50)Google Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of SurgeryRutgers-Robert Wood Johnson Medical SchoolNew BrunswickUSA

Personalised recommendations