Abstract
Acanthamoeba castellanii (JBM) was cultivated in a peptone-yeast extract-glucose medium in Erlenmeyer flasks on a rotary shaker at 30 C. Under these conditions the amebae have a generation time of 6 hr and attain maximal populations of over 3.0 X 107 organisms/ml. A decrease in dry weight and variance in biuret values was observed for this organism during exponential growth. The successful axenic cultivation of certain free-living protozoa including Tetrahymena and Paramecium has contributed greatly to the usefulness of these organisms in many biochemical and biophysical investigations. The axenic growth of Acanthamoeba (Hartmannella) spp. (Neff, 1957), soil amebae which are potentially pathogenic for man and lower primates (Culbertson et al., 1959; Butt, 1964; Fowler and Carter, 1965; Callicott, 1968), has made possible the use of these protozoa in a number of metabolic (Band, 1959; Korn, 1964; Adam and Blewett, 1967; Griffiths and Hughes, 1969) and electron microscopic studies (Bowers and Korn, 1968, 1969). However, rather long generation times and morphological heterogeneity still limit the number and type of experimental procedures which can be carried out. In this paper we present information on the axenic cultivation and growth characteristics of Acanthamoeba castellanii (JBM), a strain of ameba which grows homogeneously with a 6-hr generation time and attains large population Received for publication 26 February 1970. * Designation given to the strain of A. castellanii isolated by Jensen, Barnes, and Meyers, 1967. t This investigation was supported by Public Health Service Grant GM 13163 from the National Institute of General Medical Sciences and Public Health Training Grant AI 00137 to the University of Kansas and Public Health General Research Support Grant FR 05323 to the University of Missouri. t Present address: Kansas City, Missouri General Hospital and Medical Center. ? Present address: Penn Valley Community College, Kansas City, Missouri. numbers of > 3.0 x 107 organisms/ml. The usefulness of this organism in the development of a model system for the study of molecular biology also is considered. MATERIALS AND METHODS
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