Abstract 568: Formation of Cardiac Fibers in Matrigel Matrix
INTRODUCTION. BD Matrigel Matrix is a basement membrane preparation extracted from the Engelbreth-Holm-Swarm mouse sarcoma, a tumor rich in extracellular matrix proteins. At room temperature, the matrigel polymerizes to produce a biologically active matrix material resembling the mammalian cellular basement membrane. Matrigel is widely used for cellular attachment, cell invasion assays and as a component of cell differentiation media. Here we report a new means for its use.
METHODS. Matrigel is employed to create in vitro cardiac fibers from rat neonatal cardiomyocytes. A semi-solid “pillow” from concentrated matrigel is overlaid with a cardiomyocyte suspension in a diluted matrigel solution. Myocytes submerged in the top matrigel layer form multicellular structures of different sizes. The underling semi-solid matrigel creates an elastic environment which serves as a mechanical load against which myocytes contract.
RESULTS. This new technique creates a net of actively contracting multicellular fibers which can be maintained in vitro for up to six weeks. Presented data illustrate the appearance and development of these fibers. Dependence of the fiber size on myocyte amount and matrigel concentration is also described.
CONCLUSIONS. This new semi-3D model of cardiac muscle offers the ease of both structural and functional monitoring since fiber thickness of 50 –300 micron makes these structures suitable for live confocal microscopy (calcium and potentiometric indicators) and gives easy assess to the epitopes for immunostaining purposes.