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Effect of the Pore Structure of an Apatite-Fiber Scaffold on the Differentiation of P19.CL6 Cells into Cardiomyocytes

We previously reported that P19.CL6 cells can be cultured in porous hydroxyapatite ceramics prepared by firing green compacts consisting of apatite fibers and spherical carbon beads (150 μm in diameter). Cells cultured for 20 days in an apatite-fiber scaffold (AFS) proliferated and differentiated into cells expressing troponin T, a cardiomyocyte-specific gene, but the expression level was insufficient to support the functional maturation of cells required for biomedical device applications. In this study, we aimed to optimize the internal AFS environment for cardiomyocytes by mixing two sizes (150-and 20-μm) of carbon beads. P19.CL6 cells were cultured in AFS materials comprising different carbon ratios in the presence of alpha-MEM with (AFS+) or without (AFS-) dimethyl sulfoxide (DMSO), and cell growth and gene expression were assessed. We found that AFS(50, 1:1 ratio) is the most suitable scaffold for the proliferation and differentiation of P19.CL6 cells and the addition of DMSO to the culture medium is necessary for differentiation into cardiomyocytes. We also assessed the culture of P19.CL6 cells in AFS in a radial-flow bioreactor for several days.