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Bone marrow stromal cells contract synchronously with cardiomyocytes in a coculture system
Shinji Tomita1, Takeshi Nakatani, Shinya Fukuhara
1Department of Regenerative Medicine, National Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka 565-8565, Japan.
Objectives:
Cell transplantation is a promising therapy for improving damaged heart function. Cardiac environmental factors are thought to be powerful differentiation inducers, but their effects are not well understood because of their in vivo nature. We simulated the cardiac environment using coculture and evaluated cardiomyogenic differentiation in bone marrow stromal cells and synchronous contraction with other cardiomyocytes.
Methods:
Experiment 1. We evaluated the labeling efficiency, intensity, and pattern of green fluorescence in the transgenic mouse expressing green fluorescent protein-derived bone marrow stromal cells (GFP-BMCs) from initial plating through 8 weeks under fluorescent microscopy. Experiment 2. GFP-BMCs (10(5) cells) were cocultured with neonatal rat cardiomyocytes (10(5) cells). We also evaluated the incorporation, myogenic differentiation, and synchronous contraction of GFP-BMCs for 1 week under the same microscopy with a digital video camera.
Results:
Experiment 1. All GFP-BMCs but red blood cells maintained green fluorescence from initial plating through 8 weeks. Experiment 2. Some GFP-BMCs were incorporated in parallel with cardiomyocytes and showed myotube-like formation on day 1. On day 2, GFP-BMCs started to contract synchronously with cardiomyocytes. GFP-BMCs formed colonies and maintained synchronous contraction on day 7.
Conclusions:
Direct cell-to-cell interaction with cardiomyocytes is essential for myogenic differentiation and synchronous contraction of bone marrow cells. This coculture is a simple tool for simulating the cardiac environment and evaluating phenotypic changes in vitro.