Synchronization of Aedes albopictus mosquito cells using hydroxyurea.
We have established conditions for use of hydroxyurea, a reversible inhibitor of DNA synthesis, to synchronize the division cycle of a continuous cell line from the mosquito, Aedes albopictus. In the range of 0.15-0.25 mM hydroxyurea, an 18 h treatment, followed by removal of the drug, results in effective synchronization. When combined with the partial synchronization that occurs within 10 h of dilution and plating, more than 80% of cells treated with hydroxyurea could be recovered in the synthesis (S) phase of the cell cycle during the 4 h period after removal of the drug. The degree of synchrony was enhanced when cells were exposed to two consecutive hydroxyurea treatments spaced 10 h apart. Synchronized cells expressed maximal levels of a reporter gene when transfected immediately after removal of hydroxyurea. This is the first description of effective chemical synchronization of an insect cell line using hydroxyurea.
Gerenday
A
A
Department of Entomology, University of Minnesota, St Paul 55108, USA.
Blauwkamp
T S
TS
Fallon
A M
AM
D000330:Aedes / Q000166:cytology*; D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D006918:Hydroxyurea / Q000494:pharmacology*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology*; D014162:Transfection
D019384:Nucleic Acid Synthesis Inhibitors; D006918:Hydroxyurea