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Three‐dimensional reconstitution of embryonic cardiomyocytes in a collagen matrix: a new heart muscle model system
Author(s) -
Eschenhagen Thomas,
Fink Christine,
Remmers Ute,
Scholz Hasso,
Wattchow Jens,
Weil Joachim,
Zimmermann Wolfram,
Dohmen Hans H.,
Schäfer Hansjörg,
Bishopric Nanette,
Wakatsuki Tetsuro,
Elson Elliot L.
Publication year - 1997
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.11.8.9240969
Subject(s) - myocyte , myofilament , cardiac muscle , extracellular matrix , anatomy , embryonic stem cell , matrix (chemical analysis) , isometric exercise , biophysics , embryonic heart , chemistry , cardiac myocyte , microbiology and biotechnology , biology , biochemistry , chromatography , gene , physiology
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.—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. Three‐dimensional reconstitution of embryonic cardiomyocytes in a collagen matrix: a new heart muscle model system. FASEB J. 11, 683–694 (1997)