摘要：

A brief account of a method for axenic cultivation of Entamoeba histolytica was published by the author in 1961. Since that time many improvements have been made in the culture media and the technical procedures for handling the amebae; these are reported in detail in the present communication. In addition, the axenic cultivation of E. histolytica-like amebae is reported here for the first time. An improved diphasic medium, particularly useful for initiation of axenic cultures, is described. The new medium, unlike its predecessor, does not require supplementation with a cell-free extract of chick embryo. A clear liquid medium for initiation, maintenance, and mass cultivation of the amebae is also described. A liter of this medium which consists of Trypticase, Panmede (ox-liver digest), glucose, cysteine, ascorbic acid, and salts supplemented with horse serum and a vitamin mixture yields 100 million to 150 million E. histolytica from an inoculum of 10 million amebae. The average yield of amebic protein under these conditions is 0.87 mg/million amebae. Lower yields are obtained with the E. histolytica-like amebae. Axenization of a given strain of amebae appears to be dependent, in part, on the composition of the medium used and on the particular trypanosomatid associate employed in the monoxenic cultures supplying amebae for axenic cultures. To date, four strains of E. histolytica (200:NIH, HB-301:NIH, F-22, HK-9) and two strains of E. histolytica-like amebae (Huff and Laredo) have been grown in axenic culture for periods ranging from 2 1/2 to 7 years.

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