Factors that influence initiation and growth of 9L rat brain gliosarcoma multicellular spheroids

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 x 10(6) 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 well 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 degrees 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.