Long-term maintenance of primary myogenic cultures on a reconstituted basement membrane.
We describe a simple technique for maintaining highly contractile long-term chicken myogenic cultures on Matrigel, a gel composed of basement membrane components extracted from the Engelbreth-Holm-Swarm mouse tumor. Cultures grown on Matrigel consist of three-dimensional multilayers of cylindrical, contracting myotubes which endure for at least 60 d without myotube detachment. A Matrigel substrate increases the initial plating efficiency but does not effect cell proliferation. Large-scale differentiation in cultures maintained on Matrigel is delayed by 1 to 2 d, compared to cultures grown on gelatin-coated dishes. Long-term maintenance on Matrigel also results in increased expression of the neonatal and adult fast myosin heavy chain isoforms. Culturing of cells on a Matrigel substrate could thus facilitate the study of later events of in vitro myogenesis.
Hartley
R S
RS
Department of Biological Structure, School of Medicine, University of Washington, Seattle 98195.
Yablonka-Reuveni
Z
Z
D000818:Animals; D000911:Antibodies, Monoclonal; D001485:Basement Membrane / Q000502:physiology*; D002448:Cell Adhesion; D002454:Cell Differentiation; D002455:Cell Division; D002642:Chick Embryo; D005455:Fluorescent Antibody Technique; D005780:Gelatin; D066298:In Vitro Techniques; D009132:Muscles / Q000166:cytology*; D009218:Myosins / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D005780:Gelatin; D009218:Myosins