Multichannel perfusion culture bioluminescence reporter system for long-term detection in living cells.
We constructed a multichannel perfusion culture system. Using this device, we developed a perfusion device to detect six samples at the same time and demonstrated the response to dexamethasone of the circadian-related promoter activity in Rat1 fibroblast cells. We could detect the sequential phase shifts of the circadian peaks that are dependent on the timing of the drug treatments. We also demonstrate a temporal dual reporter assay using two kinds of secreted luciferase in the perfusion culture. The combination of secreted luciferase and multiple perfusion culture assay system reveals the effects of transient drug treatment for the pharmacological assay.
Watanabe
Toshiyuki
T
ATTO, Bunkyo-ku, Tokyo 113-0034, Japan.
Enomoto
Toshiteru
T
Takahashi
Masayuki
M
Honma
Sato
S
Honma
Ken-Ichi
K
Ohmiya
Yoshihiro
Y
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002460:Cell Line; D002940:Circadian Rhythm / Q000187:drug effects*; D003907:Dexamethasone / Q000494:pharmacology*; D004867:Equipment Design; D005347:Fibroblasts / Q000187:drug effects*; D008156:Luciferases / Q000378:metabolism*; D049408:Luminescent Agents / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods; D010477:Perfusion / Q000295:instrumentation; D011401:Promoter Regions, Genetic / Q000187:drug effects*
D049408:Luminescent Agents; C485195:luciferase, cypridina; D003907:Dexamethasone; D008156:Luciferases