Skip to main content

Microstructured Cocultures of Cardiac Myocytes and Fibroblasts: A Two-Dimensional In Vitro Model of Cardiac Tissue

  • Patrizia Camelliti (a1), Andrew D. McCulloch (a2) and Peter Kohl (a1)

Cardiac myocytes and fibroblasts are essential elements of myocardial tissue structure and function. In vivo, myocytes constitute the majority of cardiac tissue volume, whereas fibroblasts dominate in numbers. In vitro, cardiac cell cultures are usually designed to exclude fibroblasts, which, because of their maintained proliferative potential, tend to overgrow the myocytes. Recent advances in microstructuring of cultures and cell growth on elastic membranes have greatly enhanced in vitro preservation of tissue properties and offer a novel platform technology for producing more in vivo-like models of myocardium. We used microfluidic techniques to grow two-dimensional structured cardiac tissue models, containing both myocytes and fibroblasts, and characterized cell morphology, distribution, and coupling using immunohistochemical techniques. In vitro findings were compared with in vivo ventricular cyto-architecture. Cardiac myocytes and fibroblasts, cultured on intersecting 30-μm-wide collagen tracks, acquire an in vivo-like phenotype. Their spatial arrangement closely resembles that observed in native tissue: Strands of highly aligned myocytes are surrounded by parallel threads of fibroblasts. In this in vitro system, fibroblasts form contacts with other fibroblasts and myocytes, which can support homogeneous and heterogeneous gap junctional coupling, as observed in vivo. We conclude that structured cocultures of cardiomyocytes and fibroblasts mimic in vivo ventricular tissue organization and provide a novel tool for in vitro research into cardiac electromechanical function.

Corresponding author
Corresponding author. E-mail:
Hide All


Adler, C.P., Ringlage, W.P., & Böhm, N. (1981). DNS-Gehalt und Zellzahl in Herz und Leber von Kindern. Pathol Res Pract 172, 2541.
Agarkova, I., Auerbach, D., Ehler, E., & Perriard, J.C. (2000). A novel marker for vertebrate embryonic heart, the EH-myomesin isoform. J Biol Chem 275, 1025610264.
Belus, A. & White, E. (2003). Streptomycin and intracellular calcium modulate the response of single guinea-pig ventricular myocytes to axial stretch. J Physiol 546, 501509.
Bhatia, S.N. (2002). Micropatterned cell cultures and cocultures. In Methods of Tissue Engineering, A. Atala, R.P. Lanza (Eds.), pp. 121129. San Diego, CA: Academic Press.
Bogoyevitch, M.A., Clerk, A., & Sugden, P.H. (1995). Activation of the mitogen-activated protein kinase cascade by pertussis toxin-sensitive and -insensitive pathways in cultured ventricular cardiomyocytes. Biochem J 309, 437443.
Bohn, W., Wiegers, W., Beuttenmuller, M., & Traub, P. (1992). Species-specific recognition patterns of monoclonal antibodies directed against vimentin. Exp Cell Res 201, 17.
Booz, G.W. & Baker, K.M. (1995). Molecular signalling mechanisms controlling growth and function of cardiac fibroblasts. Cardiovasc Res 30, 537543.
Camelliti, P., Borg, T.K., & Kohl, P. (2005). Structural and functional characterisation of cardiac fibroblasts. Cardiovasc Res 65, 4051.
Camelliti, P., Devlin, G.P., Matthews, K.G., Kohl, P., & Green, C.R. (2004a). Spatially and temporally distinct expression of fibroblast connexins after sheep ventricular infarction. Cardiovasc Res 62, 415425.
Camelliti, P., Green, C.R., LeGrice, I., & Kohl, P. (2004b). Fibroblast network in rabbit sinoatrial node: Structural and functional identification of homogeneous and heterogeneous cell coupling. Circ Res 94, 828835.
Davies, M.J. & Pomerance, A. (1972). Quantitative study of ageing changes in the human sinoatrial node and internodal tracts. Br Heart J 34, 150152.
Gaudesius, G., Miragoli, M., Thomas, S.P., & Rohr, S. (2003). Coupling of cardiac electrical activity over extended distances by fibroblasts of cardiac origin. Circ Res 93, 421428.
Gopalan, S.M., Flaim, C., Bhatia, S.N., Hoshijima, M., Knoell, R., Chien, K.R., Omens, J.H., & McCulloch, A.D. (2003). Anisotropic stretch-induced hypertrophy in neonatal ventricular myocytes micropatterned on deformable elastomers. Biotechnol Bioeng 81, 578587.
Goshima, K. (1970). Formation of nexuses and electronic transmission between myocardial and FL cells in monolayer culture. Exp Cell Res 63, 124130.
Goshima, K. & Tonomura, Y. (1969). Synchronized beating of embryonic mouse myocardial cells mediated by FL cells in monolayer culture. Exp Cell Res 56, 387392.
Grove, B.K., Kurer, V., Lehner, C., Doetschman, T.C., Perriard, J.C., & Eppenberger, H.M. (1984). A new 185,000–dalton skeletal muscle protein detected by monoclonal antibodies. J Cell Biol 98, 518524.
Kanter, H.L., Beyer, E.C., & Saffitz, J.E. (1995). Structural and molecular determinants of intercellular coupling in cardiac myocytes. Microsc Res Tech 31, 357363.
Kohl, P. (2003). Heterogeneous cell coupling in the heart: An electrophysiological role for fibroblasts. Circ Res 93, 381383.
Kohl, P., Hunter, P., & Noble, D. (1999). Stretch-induced changes in heart rate and rhythm: Clinical observations, experiments and mathematical models. Prog Biophys Mol Biol 71, 91138.
Kohl, P., Kamkin, A.G., Kiseleva, I.S., & Noble, D. (1994). Mechanosensitive fibroblasts in the sino-atrial node region of rat heart: Interaction with cardiomyocytes and possible role. Exp Physiol 79, 943956.
Lee, A.A., Delhaas, T., Waldman, L.K., MacKenna, D.A., Villarreal, F.J., & McCulloch, A.D. (1996). An equibiaxial strain system for cultured cells. Am J Physiol 271, C1400C1408.
LeGrice, I.J., Smaill, B.H., Chai, L.Z., Edgar, S.G., Gavin, J.B., & Hunter, P.J. (1995). Laminar structure of the heart: Ventricular myocyte arrangement and connective tissue architecture in the dog. Am J Physiol 269, H571H582.
Long, C.S. & Brown, R.D. (2002). The cardiac fibroblast, another therapeutic target for mending the broken heart? J Mol Cell Cardiol 34, 12731278.
MacKenna, D., Summerour, S.R., & Villarreal, F.J. (2000). Role of mechanical factors in modulating cardiac fibroblast function and extracellular matrix synthesis. Cardiovasc Res 46, 257263.
McDevitt, T.C., Angello, J.C., Whitney, M.L., Reinecke, H., Hauschka, S.D., Murry, C.E., & Stayton, P.S. (2002). In vitro generation of differentiated cardiac myofibers on micropatterned laminin surfaces. J Biomed Mater Res 60, 472479.
Motlagh, D., Hartman, T.J., Desai, T.A., & Russell, B. (2003a). Microfabricated grooves recapitulate neonatal myocyte connexin43 and N-cadherin expression and localization. J Biomed Mater Res 67A, 148157.
Motlagh, D., Senyo, S.E., Desai, T.A., & Russell, B. (2003b). Microtextured substrata alter gene expression, protein localization and the shape of cardiac myocytes. Biomaterials 24, 24632476.
Osborn, M., Debus, E., & Weber, K. (1984). Monoclonal antibodies specific for vimentin. Eur J Cell Biol 34, 137143.
Rohr, S., Fluckiger-Labrada, R., & Kucera, J.P. (2003). Photolithographically defined deposition of attachment factors as a versatile method for patterning the growth of different cell types in culture. Pflugers Arch 446, 125132.
Rohr, S., Scholly, D.M., & Kleber, A.G. (1991). Patterned growth of neonatal rat heart cells in culture. Morphological and electrophysiological characterization. Circ Res 68, 114130.
Rook, M.B., Jongsma, H.J., & De Jonge, B. (1989). Single channel currents of homo- and heterologous gap junctions between cardiac fibroblasts and myocytes. Pflugers Arch 414, 9598.
Rook, M.B., van Ginneken, A.C.G., De Jonge, B., El Aoumari, A., Gros, D., & Jongsma, H.J. (1992). Differences in gap junction channels between cardiac myocytes, fibroblasts, and heterologous pairs. Am J Physiol 263, C959C977.
Ruwhof, C. & van der Laarse, A. (2000). Mechanical stress-induced cardiac hypertrophy: Mechanisms and signal transduction pathways. Cardiovasc Res 47, 2337.
Severs, N.J. (1995). Cardiac muscle cell interaction: From microanatomy to the molecular make-up of the gap junction. Histol Histopathol 10, 481501.
Shiraishi, I., Takamatsu, T., Minamikawa, T., Onouchi, Z., & Fujita, S. (1992). Quantitative histological analysis of the human sinoatrial node during growth and aging. Circulation 85, 21762184.
Singhvi, R., Kumar, A., Lopez, G.P., Stephanopoulos, G.N., Wang, D.I., Whitesides, G.M., & Ingber, D.E. (1994). Engineering cell shape and function. Science 264, 696698.
Sommer, J.R. & Scherer, B. (1985). Geometry of cell and bundle appositions in cardiac muscle: Light microscopy. Am J Physiol 248, H792H803.
Sun, Y., Kiani, M.F., Postlethwaite, A.E., & Weber, K.T. (2002). Infarct scar as living tissue. Basic Res Cardiol 97, 343347.
Sun, Y. & Weber, K.T. (2000). Infarct scar: A dynamic tissue. Cardiovasc Res 46, 250256.
Vliegen, H.W., van der Laarse, A., Cornelisse, C.J., & Eulderink, F. (1991). Myocardial changes in pressure overload-induced left ventricular hypertrophy. A study on tissue composition, polyploidization and multinucleation. Eur Heart J 12, 488494.
Wang, T.L., Tseng, Y.Z., & Chang, H. (2000). Regulation of connexin 43 gene expression by cyclical mechanical stretch in neonatal rat cardiomyocytes. Biochem Biophys Res Commun 267, 551557.
Whittaker, P. (1995). Unravelling the mysteries of collagen and cicatrix after myocardial infarction. Cardiovasc Res 29, 758762.
Wright, A.R. & Rees, S.A. (1997). Targeting ischaemia—Cell swelling and drug efficacy. Trends Pharmacol Sci 18, 224228.
Yamazaki, T., Komuro, I., & Yazaki, Y. (1995). Molecular mechanism of cardiac cellular hypertrophy by mechanical stress. J Mol Cell Cardiol 27, 133140.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Microscopy and Microanalysis
  • ISSN: 1431-9276
  • EISSN: 1435-8115
  • URL: /core/journals/microscopy-and-microanalysis
Please enter your name
Please enter a valid email address
Who would you like to send this to? *



Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed