Growth kinetics and histological evaluation of C6 glioma spheroid with non-adhesive culture plate.

Although C6 glioma spheroids have been widely utilized for assessment of new therapeutic modalities, they need to be generated for long-term use and special instruments. Therefore we generated a multicellular mass (spheroid) from C6 glioma cells on non-adhesive culture plates. Cells cultured in a spheroid plate (Sumiron Celltight Spheroid) spontaneously aggregated and formed a cell mass after 24 h in culture. After three days of culture, the cell mass became spherical with a diameter ranging from 300 to 650 microm. The size of the spheroid depended on the number of cells plated in each well. Histological examination indicated no spheroid had aggregations of cell death during the 8 days of culture. In the case where 1.0 x 10(3) cells were plated in each well, the size of the spheroid increased gradually and stagnated at approximately 850 microm. The present study demonstrated that a spheroid was easily generated from C6 glioma cells on a non-adhesive plate. This method could provide a new tool for analyzing therapeutic modalities for glioma.