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Starting from an optimized method to disintegrate planarian tissues and prepare purified neoblast fractions, different media and additives were tried. Hyposmotic media and layers of extracellular matrix components enhanced the adhesion of neoblasts and favoured the formation of transient processes. Proteins in the medium supported long-term survival of neoblasts that retained a spherical shape. Eventually, an isosmotic medium was devised that supported the survival of neoblasts with a viability of 46% on day 31 of primary cultures. With light microscopical techniques, no signs of differentiation were observed in these cultures. Mitoses were detected until the second day of cultivation. In contrast, cultures of total cells still displayed mitoses after 7 days of cultivation. 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