Three-dimensional reconstitution of embryonic cardiomyocytes in a collagen matrix: A new heart muscle model system

Thomas Eschenhagen, Christine Fink, Ute Remmers, Hasso Scholz, Jens Wattchow, Joachim Weil, Wolfram Zimmermann, Hans H. Dohmen, Hansjörg Schäfer, Nanette Bishopric, Tetsuro Wakatsuki, Elliot L. Elson

Research output: Contribution to journalArticle

415 Scopus citations

Abstract

A method has been developed for culturing cardiac myocytes in a collagen matrix to produce a coherently contracting 3-dimensional model heart tissue that allows direct measurement of isometric contractile force. Embryonic chick cardiomyocytes were mixed with collagen solution and allowed to gel between two Velcro-coated glass tubes. During culture, the cardiomyocytes formed spontaneously beating cardiac myocyte-populated matrices (CMPMs) anchored at opposite ends to the Velcro-covered tubes through which they could be attached to a force measuring system. Immunohistochemistry and electron microscopy revealed a highly organized tissue-like structure of α- actin and α-tropomyosin-positive cardiac myocytes exhibiting typical cross- striation, sarcomeric myofilaments, intercalated discs, desmosomes, and tight junctions. Force measurements of paced or unpaced CMPMs were performed in organ baths after 6-11 days of cultivation and were stable for up to 24 h. Force increased with frequency between 0.8 and 2.0 Hz (positive 'staircase'), increasing rest length (Starling mechanism), and increasing extracellular calcium. The utility of this system as a test bed for genetic manipulation was demonstrated by infecting the CMPMs with a recombinant β-galactosidase- carrying adenovirus. Transduction efficiency increased from about 5% (MOI 0.1) to about 50% (MOI 100). CMPMs display more physiological characteristics of intact heart tissue than monolayer cultures. This approach, simpler and faster than generation of transgenic animals, should allow functional consequences of genetic or pharmacological manipulation of cardiomyocytes in vitro to be studied under highly controlled conditions.

Original languageEnglish (US)
Pages (from-to)683-694
Number of pages12
JournalFASEB Journal
Volume11
Issue number8
DOIs
StatePublished - Jul 1997

Keywords

  • Adenovirus
  • Cell culture
  • Heart
  • Reconstituted tissue
  • Transgenic model

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

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    Eschenhagen, T., Fink, C., Remmers, U., Scholz, H., Wattchow, J., Weil, J., Zimmermann, W., Dohmen, H. H., Schäfer, H., Bishopric, N., Wakatsuki, T., & Elson, E. L. (1997). Three-dimensional reconstitution of embryonic cardiomyocytes in a collagen matrix: A new heart muscle model system. FASEB Journal, 11(8), 683-694. https://doi.org/10.1096/fasebj.11.8.9240969