title: Pokeweed mitogen stimulates DNA synthesis in cultured amoebocytes of Carcinoscorpius rotundicauda.
abstract: Cultures of Carcinoscorpius rotundicauda amoebocytes in Grace's insect (modified) medium supplemented with 20% foetal bovine serum, were stimulated with pokeweed mitogen at 0.5 to 50 micrograms/ml for 24 to 72 h at 26 degrees C. DNA synthesis was monitored by measuring the uptake of 3H-thymidine. The optimal concentration of pokeweed mitogen was established at 2.5 micrograms/ml. The ideal duration of stimulation was 48 h. Under these conditions, amoebocyte cultures showed increases in uptake of 3H-thymidine up to 55-fold. Attempts at stimulation of these amoebocytes with phytohaemagglutinin, concanavalin A and endotoxin showed no increase in 3H-thymidine uptake. Nevertheless, positive stimulation by pokeweed mitogen demonstrates the potential of such an approach to establishing a permanent amoebocyte cell line for lysate production. This stimulation was reversed by 5 mM hydroxyurea, suggesting a true induction of DNA synthesis.
authors.affiliation: Department of Zoology, National University of Singapore.
authors.forename: J L
authors.initials: JL
authors.lastname: Ding
authors.forename: J C
authors.initials: JC
authors.lastname: Kim
authors.forename: B
authors.initials: B
authors.lastname: Ho
mesh_terms: D000656:Amoeba / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D002455:Cell Division; D002460:Cell Line; D003208:Concanavalin A / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis*; D004731:Endotoxins / Q000494:pharmacology; D006737:Horseshoe Crabs / Q000378:metabolism*; D009994:Osmolar Concentration; D010835:Phytohemagglutinins / Q000494:pharmacology; D011043:Pokeweed Mitogens / Q000494:pharmacology*; D013936:Thymidine / Q000493:pharmacokinetics; D014170:Transformation, Genetic / Q000187:drug effects
chemical_list: D004731:Endotoxins; D010835:Phytohemagglutinins; D011043:Pokeweed Mitogens; D003208:Concanavalin A; D004247:DNA; D013936:Thymidine