Hostname: page-component-7d684dbfc8-4nnqn Total loading time: 0 Render date: 2023-09-25T13:05:54.468Z Has data issue: false Feature Flags: { "corePageComponentGetUserInfoFromSharedSession": true, "coreDisableEcommerce": false, "coreDisableSocialShare": false, "coreDisableEcommerceForArticlePurchase": false, "coreDisableEcommerceForBookPurchase": false, "coreDisableEcommerceForElementPurchase": false, "coreUseNewShare": true, "useRatesEcommerce": true } hasContentIssue false

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

Published online by Cambridge University Press:  12 May 2005

Patrizia Camelliti
University Laboratory of Physiology, Oxford, OX1 3PT, UK
Andrew D. McCulloch
Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093-0412, USA
Peter Kohl
University Laboratory of Physiology, Oxford, OX1 3PT, UK
Get access


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.

Research Article
© 2005 Microscopy Society of America

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)



Adler, C.P., Ringlage, W.P., & Böhm, N. (1981). DNS-Gehalt und Zellzahl in Herz und Leber von Kindern. Pathol Res Pract 172, 2541.Google Scholar
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.Google Scholar
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.Google Scholar
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.Google Scholar
Bohn, W., Wiegers, W., Beuttenmuller, M., & Traub, P. (1992). Species-specific recognition patterns of monoclonal antibodies directed against vimentin. Exp Cell Res 201, 17.Google Scholar
Booz, G.W. & Baker, K.M. (1995). Molecular signalling mechanisms controlling growth and function of cardiac fibroblasts. Cardiovasc Res 30, 537543.Google Scholar
Camelliti, P., Borg, T.K., & Kohl, P. (2005). Structural and functional characterisation of cardiac fibroblasts. Cardiovasc Res 65, 4051.Google Scholar
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.Google Scholar
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.Google Scholar
Davies, M.J. & Pomerance, A. (1972). Quantitative study of ageing changes in the human sinoatrial node and internodal tracts. Br Heart J 34, 150152.Google Scholar
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.Google Scholar
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.Google Scholar
Goshima, K. (1970). Formation of nexuses and electronic transmission between myocardial and FL cells in monolayer culture. Exp Cell Res 63, 124130.Google Scholar
Goshima, K. & Tonomura, Y. (1969). Synchronized beating of embryonic mouse myocardial cells mediated by FL cells in monolayer culture. Exp Cell Res 56, 387392.Google Scholar
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.Google Scholar
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.Google Scholar
Kohl, P. (2003). Heterogeneous cell coupling in the heart: An electrophysiological role for fibroblasts. Circ Res 93, 381383.Google Scholar
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.Google Scholar
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.Google Scholar
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.Google Scholar
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.Google Scholar
Long, C.S. & Brown, R.D. (2002). The cardiac fibroblast, another therapeutic target for mending the broken heart? J Mol Cell Cardiol 34, 12731278.Google Scholar
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.Google Scholar
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.Google Scholar
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.Google Scholar
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.Google Scholar
Osborn, M., Debus, E., & Weber, K. (1984). Monoclonal antibodies specific for vimentin. Eur J Cell Biol 34, 137143.Google Scholar
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.Google Scholar
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.Google Scholar
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.Google Scholar
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.Google Scholar
Ruwhof, C. & van der Laarse, A. (2000). Mechanical stress-induced cardiac hypertrophy: Mechanisms and signal transduction pathways. Cardiovasc Res 47, 2337.Google Scholar
Severs, N.J. (1995). Cardiac muscle cell interaction: From microanatomy to the molecular make-up of the gap junction. Histol Histopathol 10, 481501.Google Scholar
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.Google Scholar
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.Google Scholar
Sommer, J.R. & Scherer, B. (1985). Geometry of cell and bundle appositions in cardiac muscle: Light microscopy. Am J Physiol 248, H792H803.Google Scholar
Sun, Y., Kiani, M.F., Postlethwaite, A.E., & Weber, K.T. (2002). Infarct scar as living tissue. Basic Res Cardiol 97, 343347.Google Scholar
Sun, Y. & Weber, K.T. (2000). Infarct scar: A dynamic tissue. Cardiovasc Res 46, 250256.Google Scholar
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.Google Scholar
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.Google Scholar
Whittaker, P. (1995). Unravelling the mysteries of collagen and cicatrix after myocardial infarction. Cardiovasc Res 29, 758762.Google Scholar
Wright, A.R. & Rees, S.A. (1997). Targeting ischaemia—Cell swelling and drug efficacy. Trends Pharmacol Sci 18, 224228.Google Scholar
Yamazaki, T., Komuro, I., & Yazaki, Y. (1995). Molecular mechanism of cardiac cellular hypertrophy by mechanical stress. J Mol Cell Cardiol 27, 133140.Google Scholar