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Functional Toxicology and Pharmacology Test of Cell Induced Mechanical Tensile Stress in 2D and 3D Tissue Cultures

  • Gerhard M. Artmann
  • Jürgen Hescheler
  • Haritha Meruvu
  • Sefa Kizildag
  • Aysegül Artmann
Chapter

Abstract

Mechanical forces/tensile stresses are critical determinants of cellular growth, differentiation and migration patterns in health and disease. The innovative “CellDrum technology” was designed for measuring mechanical tensile stress of cultured cell monolayers/thin tissue constructs routinely. These are cultivated on very thin silicone membranes in the so-called CellDrum. The cell layers adhere firmly to the membrane and thus transmit the cell forces generated. A CellDrum consists of a cylinder which is sealed from below with a 4 μm thick, biocompatible, functionalized silicone membrane. The weight of cell culture medium bulbs the membrane out downwards. Membrane indentation is measured. When cells contract due to drug action, membrane, cells and medium are lifted upwards. The induced indentation changes allow for lateral drug induced mechanical tension quantification of the micro-tissues. With hiPS-induced (human) Cardiomyocytes (CM) the CellDrum opens new perspectives of individualized cardiac drug testing. Here, monolayers of self-beating hiPS-CMs were grown in CellDrums. Rhythmic contractions of the hiPS-cells induce membrane up-and-down deflections. The recorded cycles allow for single beat amplitude, single beat duration, integration of the single beat amplitude over the beat time and frequency analysis. Dose effects of agonists and antagonists acting on Ca2+ channels were sensitively and highly reproducibly observed. Data were consistent with published reference data as far as they were available. The combination of the CellDrum technology with hiPS-Cardiomyocytes offers a fast, facile and precise system for pharmacological and toxicological studies. It allows new preclinical basic as well as applied research in pharmacolgy and toxicology.

Notes

Acknowledgements

Some of our teachers remain a role model for our entire life. Gerhard Artmann’s role models remain the professors Y. C. Fung, Shu Chien (both UCSD, San Diego, USA), Georg Büldt (Research Center Juelich, Germany) and Peter Paufler, physicist and crystallographer (University of Dresden, Germany). For providing stem cell expertise and free disposal of hIPS derived, highly purified and well characterized hIPS derived cardiomyocytes as well as for his excellent scientific advice and kindness we owe special thanks to Professor Jürgen Hescheler.

All technological developments and scientific tasks related to the CellDrum technology were carried out in the laboratories and under the scientific direction of Professors Gerhard M. as well as Aysegül Artmann. Significant contributions to the work were made by Professor J. Trzewik our former Ph.D. student and major CellDrum co-inventor. Working with him was scientifically and technologically very productive. Thank you, Juergen. Professors Artmann thank their doctoral candidate M. Gossmann for his lab work. As his teachers, however, they are thoughtfully looking back at the time with him. Another but extremely talented doctoral candidate was P. Linder. He worked for more than a decade and a half intensively with us. We thank the young Prof. Dr. Ilya Digel for the excellent cooperation in the Institute of Bioengineering. In particular, we thank him for many stimulating scientific discussions and his ongoing loyalty.

Quite a number of CellDrum projects were financed by numerous grants to Professors Artmann received by the State of North Rhine Westphalia (NRW) as well as by funding institutions of the Government of the Federal Republic of Germany (BMBF, BMWI).

References

  1. 1.
    Anfinsen, C. B. (1973, July 20). Principles that govern the folding of protein chains. Science, 181(4096), 223–230.CrossRefGoogle Scholar
  2. 2.
    Artmann, G. M., Digel, I., Zerlin, K. F., Maggakis-Kelemen, C., Linder, P., Porst, D., et al. (2009). Hemoglobin senses body temperature. European Biophysics Journal, 38(5), 589–600.CrossRefGoogle Scholar
  3. 3.
    Artmann, G. M. (2000, August). Device and method for the measurement of forces from living materials. Patents, Ref. No: US020040033482A1, 08/2000, AU000007638401A, 08/2000, EP000001311850B1, 08/2000, CA 2420141, 07/2002.Google Scholar
  4. 4.
  5. 5.
    Artmann, G. M. (2015, November). 6. Deutscher Querdenker Kongreß, Award Winner: Category—”Pioneering”.Google Scholar
  6. 6.
    Artmann, G. M., & Chien, S. (2008). Bioengineering in cell and tissue research. Berlin, Heidelberg: Springer.Google Scholar
  7. 7.
    Artmann, G. M., Burns, L., Canaves, J. M., Temiz-Artmann, A., Schmid-Schönbein, G. W., Chien, S., et al. (2004). Circular dichroism spectra of human hemoglobin reveal a reversible structural transition at body temperature. European Biophysics Journal, 33(6), 490–496.Google Scholar
  8. 8.
    Artmann, G. M., Kelemen, C., Porst, D., Büldt, G., & Chien, S. (1998). Temperature transitions of protein properties in human red blood cells. Biophysical Journal, 75(6), 3179–3183.CrossRefGoogle Scholar
  9. 9.
  10. 10.
    Bell, R. M., Mocanu, M. M., & Yellon, D. M. (2011, June). Retrograde heart perfusion: The Langendorff technique of isolated heart perfusion. Journal of Molecular and Cellular Cardiology, 50(6), 940–950.CrossRefGoogle Scholar
  11. 11.
    Bissell, M. J., Hall, H. G., & Parry, G. (1982, November 7). How does the extracellular matrix direct gene expression? Journal of Theoretical Biology, 1(7), 31–68.CrossRefGoogle Scholar
  12. 12.
    Bootman, M. D., Higazi, D. R., Coombes, S., & Roderick, H. L. (2006). Calcium signalling during excitation-contraction coupling in mammalian atrial myocytes. Journal of Cell Science, 119, 3915–3925.CrossRefGoogle Scholar
  13. 13.
    Brown, R. A., Prajapati, R., McGrouther, D. A., Yannas, I. V., & Eastwood, M. (1998, June). Tensional homeostasis in dermal fibroblasts: Mechanical responses to mechanical loading in three-dimensional substrates. Journal of Cellular Physiology, 323–332.CrossRefGoogle Scholar
  14. 14.
    Burridge, P. W., Li, Y. F., Matsa, E., Wu, H., Ong, S., Sharma, A., et al. (2016). Human induced pluripotent stem cell-derived cardiomyocytes recapitulate the predilection of breast cancer patients to doxorubicin-induced cardiotoxicity. Nature Medicine, 22(5), 547–556.CrossRefGoogle Scholar
  15. 15.
    Burton, K., & Taylor, D. L. (1997, January 30). Traction forces of cytokinesis measured with optically modified elastic substrata. Nature, 385, 450–454.CrossRefGoogle Scholar
  16. 16.
    Campàs, O., Mammoto, T., Hasso, S., Sperling, R. A., O’Connell, D., Bischof, A. G., et al. (2014). Quantifying cell-generated mechanical forces within living embryonic tissues. Nature Methods, 11, 183–189.CrossRefGoogle Scholar
  17. 17.
    Cerbai, E., Crucitti, A., Sartiani, L., De Paoli, P., Pino, R., Rodriguez, M. L., et al. (2000, January). Long-term treatment of spontaneously hypertensive rats with Losartan and electrophysiological remodeling of cardiomyocytes. Cardiovascular Research, 45(2), 388–396.CrossRefGoogle Scholar
  18. 18.
    Chen, C. S., Alonso, J. L., Ostuni, E., Whitesides, G. M., & Ingber, D. E. (2003, July 25). Cell shape provides global control of focal adhesion assembly. Biochemical and Biophysical Research Communications, 307(2), 355–361.CrossRefGoogle Scholar
  19. 19.
    Chien, S. (2007, March). Mechanotransduction and endothelial cell homeostasis: The wisdom of the cell. American Journal of Physiology—Heart and Circulatory Physiology, 292(3), H1209–H1224.CrossRefGoogle Scholar
  20. 20.
    Chien, S., Sung, K. L., Skalak, S., Usami, A., & Tözeren, A. (1978, November). Theoretical and experimental studies on viscoelastic properties of erythrocyte membrane. Biophysical Journal, 2, 463–487.CrossRefGoogle Scholar
  21. 21.
    Cost, A. L., Ringer, P., Grashoff, A. C., & Grashoff, C. (2015). How to measure molecular forces in cells: A guide to evaluating genetically-encoded FRET-based tension sensors. Cellular and Molecular Bioengineering, 8(1), 96–105.CrossRefGoogle Scholar
  22. 22.
    David, B. (2000, May 4). A surprising simplicity to protein folding. Nature, 405, 39–42.CrossRefGoogle Scholar
  23. 23.
    Devalla, H. D., Schwach, V., Ford, J. W., Milnes, J. T., Haou, S., Jackson, C., et al. (2015). Atrial-like cardiomyocytes from human pluripotent stem cells are a robust preclinical model for assessing atrial-selective pharmacology. EMBO Molecular Medicine, 7, 394–410.CrossRefGoogle Scholar
  24. 24.
    Dumont, S., & Prakash, M. (2014, November 5). Emergent mechanics of biological structures. Molecular Biology of the Cell, 25(22), 3461–3465.CrossRefGoogle Scholar
  25. 25.
    Eastwood, M., McGrouther, D., & Brown, R. A. (1994, November 11). A culture force monitor for measurement of contraction forces generated in human dermal fibroblast cultures: Evidence for cell-matrix mechanical signalling. Biochimica et Biophysica Acta, 1201(2), 186–192.CrossRefGoogle Scholar
  26. 26.
    Eder, A., Vollert, I., Hansen, A., & Eschenhagen, T. (2016, January 15). Human engineered heart tissue as a model system for drug testing. Advanced Drug Delivery Reviews, 96, 214–224.CrossRefGoogle Scholar
  27. 27.
    Fung, Y. C. (1993). Biomechanics: Mechanical properties of living tissuses. New York: Springer.CrossRefGoogle Scholar
  28. 28.
    Gawain, T., Burnham, N. A., Camesano, T. A., & Wen, Q. (2013). Measuring the mechanical properties of living cells using atomic force microscopy. Journal of Visualized Experiments, 76, 50497.Google Scholar
  29. 29.
    Gayrard, C., & Borghi, N. (2016). FRET-based molecular tension microscopy. Methods, 94, 33–42.CrossRefGoogle Scholar
  30. 30.
    Giancotti, F. G., & Ruoslahti, E. (1999, August 13). Integrin signaling. Science, 285(5430), 1028–1033.CrossRefGoogle Scholar
  31. 31.
    Goßmann, M., Frotscher, R., Linder, P., Neumann, S., Bayer, R., Epple, M., et al. (2016). Mechano-pharmacological characterization of cardiomyocytes derived from human induced pluripotent stem cells. Cellular Physiology and Biochemistry, 38, 1182–1198.CrossRefGoogle Scholar
  32. 32.
    Hamel, V., Cheng, K., Liao, S., Lu, A., Zheng, Y., Chen, Y., et al. (2017, February 20). De novo human cardiomyocytes for medical research: Promises and challenges. Stem Cells International, 2017, 4528941.  https://doi.org/10.1155/2017/4528941.
  33. 33.
    Harris, A. K., Wild, P., & Stopak, D. (1980, April 11). Silicone rubber substrata: A new wrinkle in the study of cell locomotion. Science, 208(4440), 177–179.CrossRefGoogle Scholar
  34. 34.
    Heath, J. P., & Dunn, G. A. (1978). Cell to substratum contacts of chick fibroblasts and their relation to the microfilament system. A correlated interference-reflexion and high-voltage electron-microscope study. Journal of Cell Science, 197–212.Google Scholar
  35. 35.
    Heisenberg, C. P. (2017). Cell biology: Stretched divisions. Nature, 543(7643), 4443–4444.CrossRefGoogle Scholar
  36. 36.
    Helenius, J., Heisenberg, C. P., Gaub, H. E., & Muller, D. J. (2008). Single-cell force spectroscopy. Journal of Cell Science, 121, 1785–1791.CrossRefGoogle Scholar
  37. 37.
    Heras-Bautista, C. O., Katsen-Globa, A., Schloerer, N. E., Dieluweit, S., Abd El Aziz, O. M., Peinkofer, G., et al. (2014). The influence of physiological matrix conditions on permanent culture of induced pluripotent stem cell-derived cardiomyocytes. Biomaterials, 35(26), 7374–7385.CrossRefGoogle Scholar
  38. 38.
    Hochmuth, R. M. (2000). Micropipette aspiration of living cells. Journal of Biomechanics, 33(15), 15–22.CrossRefGoogle Scholar
  39. 39.
    Holmgren, G., Synnergren, J., Bogestål, Y., Améen, C., Åkesson, K., Holmgren, S., et al. (2015). Identification of novel biomarkers for doxorubicin-induced toxicity in human cardiomyocytes derived from pluripotent stem cells. Toxicology, 328, 102–111.CrossRefGoogle Scholar
  40. 40.
    Horn, M. A., Graham, H. K., Richards, M. A., Clarke, J. D., Greensmith, D. J., Briston, S. J., et al. (2012, July). Age-related divergent remodeling of the cardiac extracellular matrix in heart failure: Collagen accumulation in the young and loss in the aged. Journal of Molecular and Cellular Cardiology, 53(1), 82–90.CrossRefGoogle Scholar
  41. 41.
    Hwang, E., Park, S.-Y., Sun, Z.-W., Shin, H.-S., Lee, D.-G., & Yi, T. H. (2014). The protective effects of fucosterol against skin damage in UVB-irradiated human dermal fibroblasts. Marine Biotechnology (NY), 16(3), 361–370.CrossRefGoogle Scholar
  42. 42.
    Ionta, V., Liang, W., Kim, E. H., Rafie, R., Giacomello, A., Marbán, E., et al. (2015, January). SHOX2 overexpression favors differentiation of embryonic stem cells into cardiac pacemaker cells, improving biological pacing ability. Stem Cell Reports, 4(1), 129–142.CrossRefGoogle Scholar
  43. 43.
    Iskratsch, T., Wolfenson, H., & Sheetz, M. P. (2014). Appreciating force and shape—The rise of mechanotransduction in cell biology. Nature Reviews Molecular Cell Biology, 15, 825–833.  https://doi.org/10.1038/nrm3903.CrossRefGoogle Scholar
  44. 44.
    Jin, T., Li, L., Siow, R. C. M., & Liu, K.-K. (2015). A novel collagen gel-based measurement technique for quantitation of cell contraction force. Journal of The Royal Society Interface, 12(106), 20141365–20141365.CrossRefGoogle Scholar
  45. 45.
    Joung, Y.-H. (2013). Development of implantable medical devices: From an engineering perspective. International Neurourology Journal 17(3), 98.CrossRefGoogle Scholar
  46. 46.
    Kapoor, N., Liang, W., Marbán, E., & Cho, H. C. (2013). Direct conversion of quiescent cardiomyocytes to pacemaker cells by expression of Tbx18. Nature Biotechnology, 31, 54–62.CrossRefGoogle Scholar
  47. 47.
    Khalili, A. A., & Ahmad, M. R. (2015). A review of cell adhesion studies for biomedical and biological applications. International Journal of Molecular Sciences, 16(8), 18149–18184.CrossRefGoogle Scholar
  48. 48.
    Kirmse, M., Otto, H., & Ludwig, T. (2011). Interdependency of cell adhesion, force generation and extracellular proteolysis in matrix remodeling. Journal of Cell Science, 124, 1857–1866.CrossRefGoogle Scholar
  49. 49.
    Kolodney, M. S., & Wysolmerski, R. B. (1992, April 1). Isometric contraction by fibroblasts and endothelial cells in tissue culture: A quantitative study. The Journal of Cell Biology, 117, 73–82.CrossRefGoogle Scholar
  50. 50.
    Korn, E. D., Carlier, M. F., & Pantaloni, D. (1987, October 30). Actin polymerization and ATP hydrolysis. Science, 238(4827), 638–644.CrossRefGoogle Scholar
  51. 51.
    Kumar, V., Abul, A. K., & Aster, J. C. (2014). Robbins & Cotran pathologic basis of disease (9th Ausg.). Philadelphia: Elsevier.Google Scholar
  52. 52.
    Kurazumi, H., Kubo, M., Ohshima, M., Yamamoto, Y., Takemoto, Y., Suzuki, R., et al. (2011). The effects of mechanical stress on the growth, differentiation, and paracrine factor production of cardiac stem cells. PLoS One, 6(12).CrossRefGoogle Scholar
  53. 53.
    Langendorff, O. (1895). Untersuchungen am überlebenden Säugetierherzen. Pflugers Arch, 61, 291 ff.CrossRefGoogle Scholar
  54. 54.
    Lanir, Y., & Fung, Y. C. (1974). Two-dimensional mechanical properties of rabbit skin. I. Experimental system. Journal of Biomechanics, 7(1), 29–34.CrossRefGoogle Scholar
  55. 55.
    Lehto, H., Talo, A., Tirri, R., & Vornanen, M. (1983, August). Membrane potential oscillations in enzymatically isolated rat myocardial cells. Acta Physiologica, 118(4), 385–391.CrossRefGoogle Scholar
  56. 56.
    Linder, P., Trzewik, J., Rüffer, M., & Artmann, G. M. (2010, January). Contractile tension and beating rates of self-exciting monolayers and 3D-tissue constructs of neonatal rat cardiomyocytes. Medical & Biological Engineering & Computing, 48, 59–65.CrossRefGoogle Scholar
  57. 57.
    Liu, K.-K., & Oyen, M. L. (2014). Nanobiomechanics of living materials. Interface Focus, 4(2), 20140001–20140001.CrossRefGoogle Scholar
  58. 58.
    Mannhardt, I., Breckwoldt, K., Letuffe-Brenière, D., Schaaf, S., Schulz, H., Neuber, C., et al. (2016, July 12). Human engineered heart tissue: Analysis of contractile force. Stem Cell Reports, 7(1), 29–42.CrossRefGoogle Scholar
  59. 59.
    Pollard, T. D. (1986). Assembly and dynamics of the actin filament system in nonmuscle cells. Journal of Cellular Biochemistry, 31(2), 87–95.CrossRefGoogle Scholar
  60. 60.
    Ringer, P., Colo, G., Fässler, R., & Grashoff, C. (2017, April 4). Sensing the mechano-chemical properties of the extracellular matrix. Matrix Biology (in press).CrossRefGoogle Scholar
  61. 61.
    Ruoslahti, E. (1997, May 30). Stretching is good for a cell. Science, 276(5317), 1345.CrossRefGoogle Scholar
  62. 62.
    Schimid, S. G., Kosawada, T., Skalak, R., & Chien, S. (1995, May 01). Membrane model of endothelial cells and leukocytes. A proposal for the origin of a cortical stress. Journal of Biomechanical Engineering, 117(2), 171–178.CrossRefGoogle Scholar
  63. 63.
    Shen, Y., Nakajima, M., Kojima, S., Homma, M., Kojima, M., & Fukuda, T. (2011, October 5). Single cell adhesion force measurement for cell viability identification using an AFM cantilever-based micro putter. Measurement Science and Technology, 22(11), 115802.CrossRefGoogle Scholar
  64. 64.
    Soiné, J. R. D., Hersch, N., Dreissen, G., Hampe, N., Hoffmann, B., Merkel, R., & Schwarz, U. S. (2016). Measuring cellular traction forces on non-planar substrates. Interface Focus, 6(5), 20160024CrossRefGoogle Scholar
  65. 65.
    Solaro, R. J. (2007, December). Mechanisms of the Frank-Starling Law of the heart: The beat goes on. Biophysics Journal, 15, 93(12), 4095–4096.Google Scholar
  66. 66.
    Stabley, D. R., Jurchenko, C., Marshall, S. S., & Salaita, K. S. (2012). Visualizing mechanical tension across membrane receptors with a fluorescent sensor. Nature Methods, 9, 64–67.CrossRefGoogle Scholar
  67. 67.
    Sugimura, K., Lenne, P. F., & Graner, F. (2016). Measuring forces and stresses in situ in living tissues. Development, 143, 186–196.CrossRefGoogle Scholar
  68. 68.
    Sutherland, F. J., Shattock, M. J., Baker, K. E., & Hearse, D. J. (2003, November). Mouse isolated perfused heart: Characteristics and cautions. Clinical and Experimental Pharmacology and Physiology, 30(11), 867–878.CrossRefGoogle Scholar
  69. 69.
    Theret, D. P., Levesque, M. J., Sato, M., Nerem, R. M., & Wheeler, L. T. (1988, August 1). The application of a homogeneous half-space model in the analysis of endothelial cell micropipette measurements. Journal of Biomechanical Engineering, 110(3), 190–199.CrossRefGoogle Scholar
  70. 70.
    Tirri, R., & Lehto, H. (1984). Alpha and beta adrenergic control of contraction force of perch heart (Perca fluviatilis) in vitro. Comparative biochemistry and physiology C, 77(2), 301–304.CrossRefGoogle Scholar
  71. 71.
    Tomasek, J. J., Gabbiani, G., Hinz, B., Chaponnier, C., & Brown, R. A. (2002, May). Myofibroblasts and mechano-regulation of connective tissue remodelling. Nature Reviews Molecular Cell Biology, 3(5), 349–363.CrossRefGoogle Scholar
  72. 72.
    Trzewik, J., Ates, M., & Artmann, G. A. (2002). A novel method to quantify mechanical tension in cell monolayers. Biomedizinische Technik/Biomedical Engineering, 47(s1a), 379–381.CrossRefGoogle Scholar
  73. 73.
  74. 74.
    Trzewik, J., Linder, P., & Zerlin, K. F. (2008). How strong is the beating of cardiac myocytes?—The cell drum solution. In Bioengineering in cell and tissue research. Berlin, Heidelberg: Springer. ISBN 978–3-540-75408.Google Scholar
  75. 75.
    Verma, D., Bajpai, V. K., Ye, N., Maneshi, M. M., Jetta, D., Andreadis, S. T., et al. (2017). Flow induced adherens junction remodeling driven by cytoskeletal forces. Experimental Cell Research, 359(2), 327–336.CrossRefGoogle Scholar
  76. 76.
    Weng, Z., Kong, C.-W., Lihuan, R., Ioannis, K., Lin, G., Jiaozi, H., et al. (2014, February 25). A simple, cost-effective but highly efficient system for deriving ventricular cardiomyocytes from human pluripotent stem Cells. Stem Cells and Development, 23(14), 1704–1716.CrossRefGoogle Scholar
  77. 77.
    Wobus, A. M., Wallukat, G., & Hescheler, J. (1991). Pluripotent mouse embryonic stem cells are able to differentiate into cardiomyocytes expressing chronotropic responses to adrenergic and cholinergic agents and Ca2+ channel blockers. Differentiation, 48(3), 173–182.CrossRefGoogle Scholar
  78. 78.
    Zigrino, P., Brinckmann, J., Niehoff, A., Lu, Y., Giebeler, N., Eckes, B., et al. (2016). Fibroblast-derived MMP-14 regulates collagen homeostasis in adult skin. Journal of Investigative Dermatology, 136(8), 1575–1583.CrossRefGoogle Scholar
  79. 79.
    Zimmermann, W. H., Eschenhagen, T. (2005, December). Engineering myocardial tissue. Circulation Research, 97(12), 1220–1231.CrossRefGoogle Scholar
  80. 80.

Copyright information

© Springer Nature Singapore Pte Ltd. 2018

Authors and Affiliations

  • Gerhard M. Artmann
    • 1
  • Jürgen Hescheler
    • 1
  • Haritha Meruvu
    • 2
  • Sefa Kizildag
    • 3
    • 4
  • Aysegül Artmann
    • 3
  1. 1.Institute of Neurophysiology, University of CologneCologneGermany
  2. 2.Department of BioengineeringGaziosmanpasa UniversityTokatTurkey
  3. 3.Institute for Bioengineering, Medical and Molecular BiologyUniversity of Applied Sciences AachenJuelichGermany
  4. 4.Department of Medical Biology, Faculty of MedicineDokuz Eylul UniversityIzmirTurkey

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