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Synchronization of Aedes albopictus mosquito cells using hydroxyurea
Author(s) -
Gerenday A.,
Blauwkamp T. Shepard,
Fallen A. M.
Publication year - 1997
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/j.1365-2583.1997.tb00087.x
Subject(s) - aedes albopictus , biology , cell cycle , dna synthesis , transfection , aedes , cell division , cell culture , cell synchronization , microbiology and biotechnology , dna , cell , botany , biochemistry , genetics , aedes aegypti , larva
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.