Chapter 2 Cultivation and Synchronous Morphogenesis of Dictyostelium under Controlled Experimental Conditions

Abstract

Publisher Summary Cell populations are cultivated for a number of purposes: (1) to maintain strains; (2) to isolate mutants, revertants, or ploidal variants from a parent stock; (3) to harvest large numbers of cells for isolation of DNA, plasma membranes, proteins, and other entities; and (4) to obtain physiologically homogeneous populations of vegetative amoebae that can then be made to proceed synchronously through part or all of the morphogenetic sequence leading to fruiting body construction. To achieve these ends, the myxamoebae are grown axenically or in association with bacteria, on solid substratum or in liquid suspension, clonally or in mass culture. Detailed protocols that cover these exigencies are given. Three methods for strain preservation generally employed: lyophilization, silica gel desiccation and frozen storage. Under optimal conditions, vegetative cell populations harvested from nutrient medium and dispensed homogeneously on a solid substratum can be made to exhibit remarkably consistent morphogenetic behavior in respect to the timing and patterns of aggregation and of fruiting body construction.