During fruiting-body construction by Dictyostelium discoideum, the formation and subsequent maintenance of the multicellular assembly involve two stage-specific cohesive systems that are acquired sequentially and are distinguishable on serological and genetic grounds. We demonstrated that both systems, termed aggregation related (AR) and postaggregation related (PAR), can function in vitro. Ghosts prepared from cells of the wild-type and of a cohesion-defective mutant that were harvested during growth and at aggregation and postaggregative stages of fruiting-body construction exhibited the same cohesive properties as the cells from which they were derived. Membrane fragments prepared from the ghosts by mechanical disruption retained these cohesive properties.