Abstract
This report details optimum conditions for initiating and growing 9L rat brain gliosarcoma multicellular spheroids. Best results were achieved when 0.5 to 2.0 × 106 cells were seeded initially into agarose (0.5 to 1.0%)-coated Petri dishes. Spheroids initiated in this manner appeared to grow in spinner culture at a rate slightly faster than spheroids initiated and grown in spinner culture. An optimum growth rate was obtained for individual spheroids grown in the wells of cluster dishes coated with 0.5 to 0.75% agarose in growth medium containing 10% newborn calf serum and 0.9 g glucose per liter.
Three methods for dissociation of spheroids at both room temperature and 37° have been developed. The “race track” method of fractional dissociation produced a constant and similar number of cells for each disaggregation interval, and the plating efficiency of cells in each fraction was similar.
This research was supported by NIH Grants CA-13525 and CA-19992 and by the Morris Stulsaft Foundation.