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 ef...

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Bibliographic Details
Published in:Insect molecular biology 1997-05, Vol.6 (2), p.191-196
Main Authors: Gerenday, A, Blauwkamp, T.S, Fallon, A.M
Format: Article
Language:English
Subjects:
Aedes - cytology
Aedes albopictus
Animals
Cell Cycle
cell division
Cell Line
cell lines
chloramphenicol acetyltransferase
Culicidae
cultured cells
DNA replication
DNA synthesis
ecdysone
estrus synchronization
Freshwater
hydroxycarbamide
hydroxyurea
Hydroxyurea - pharmacology
Keywords
mosquito
Nucleic Acid Synthesis Inhibitors - pharmacology
reporter genes
Transfection
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Summary: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 cella 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.
ISSN:0962-1075
1365-2583
DOI:10.1111/j.1365-2583.1997.tb00087.x