Abstract
Multicellular spheroids are obtained in a variety of three-dimensional (3D) culture systems without the use of supporting scaffold. We present here a 3D culture method that resulted in a multicellular sheet under scaffold-free conditions. A floating disk-shaped 3D culture was prepared by magnetic levitation of B16F10 cells that has ingested Fe3O4-containing fibroin microspheres. The melanoma disk grew up to 19 mm in diameter and the thickness was ranged between 80 and 100 μm. The 3D culture was filled with closely packed cells that were proliferating exponentially at a specific growth rate of µ = 0.015 h-1. Approximately half of the cells were Ki-67 positive with no detectable levels of apoptotic or autophagic cells. However, the percentage of propidium iodide-permeable cells was 8.5 ± 1.2 %, which was probably due to physical damage in the cell membrane caused by Fe3O4-containing microspheres under a strong magnetic field. Melanin production increased by a factor of 3.0-3.7 in the 3D culture, due to an increased population of pigmented cells. This study presented a surface 3D culture of B16F10 cells without the use of a scaffold based on magnetic levitation.