Clonal growth of Entamoeba in agar: some applications of this technique to the study of their cell biology.
A new technique for growing single Entamoeba trophozoites into colonies in agar has been developed. This method depended upon axenic cultivation and utilized Diamond's new TYI-S-33 medium. The present paper describes several different types of experiment which have utilized the agar technique. a) Cloning. The isolation of clones from agar way easy and successful. Clones of the HM-1 strain did not differ in virulence for newborn hamster liver or in colony morphology or colony forming efficiency. b) Viability measurements. Colony forming efficiency (CFE) was proportional to the number of viable cells in a culture. Pilot studies with the amebacides, metronidazole and emetine, showed that the agar method should be useful in drug testing. Colchicine at high concentration inhibited clonal growth in a non-specific manner. c) E. histolytica HM-1 and E. invadens cells were rapidly killed by exposure to 42 degree C but survived relatively well at 0 degree C. d) Hemolysis. In a preliminary experiment HM-1 colonies did not produce halo's of hemolysis when grown in agar containing sheep red blood cells.
D000362:Agar; D000818:Animals; D002999:Clone Cells / Q000187:drug effects; D003078:Colchicine / Q000494:pharmacology; D003080:Cold Temperature; D003115:Colony-Stimulating Factors / Q000032:analysis; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods*; D004640:Emetine / Q000494:pharmacology; D004748:Entamoeba histolytica / Q000166:cytology / Q000187:drug effects / Q000254:growth & development*; D006358:Hot Temperature; D008795:Metronidazole / Q000494:pharmacology; D010274:Parasitology / Q000379:methods*; D010865:Pilot Projects; D013997:Time Factors
D003115:Colony-Stimulating Factors; D003470:Culture Media; D008795:Metronidazole; D000362:Agar; D003078:Colchicine; D004640:Emetine