TY - JOUR
T1 - Real-time monitoring of reactive oxygen species production during differentiation of human monocytic cell lines (THP-1)
AU - Kasai, Shigenobu
AU - Shiku, Hitoshi
AU - Torisawa, Yu Suke
AU - Noda, Hiroyuki
AU - Yoshitake, Jun
AU - Shiraishi, Takuo
AU - Yasukawa, Tomoyuki
AU - Watanabe, Toshiaki
AU - Matsue, Tomokazu
AU - Yoshimura, Tetsuhiko
N1 - Funding Information:
This work was supported in part by a Grant-in-Aid for Scientific Research from the Japan Society for Promotion of Science (17550146 (T.Y.)) and for Scientific Research on Priority Areas from the Ministry of Education, Culture, Sports, Science and Technology (15087212, 17710112), Japan.
PY - 2005/9/6
Y1 - 2005/9/6
N2 - Human monocytic leukemia cell line THP-1 differentiates into macrophages by phorbol myristate acetate (PMA) treatment. We report real-time monitoring of reactive oxygen species (ROS) production during the differentiation process. The production of ROS by THP-1 with several hours time scale has been detected using electrochemical and chemiluminescence methods. The increase in oxidation current derived from ROS arising from THP-1 was observed between 1 and 10 h after the commencement of treatment with 20 nM PMA. The response was unusually slow compared to the quick oxidative response observed in the macrophages. Chemiluminescence methods were used to further investigate the ROS production rate by carrying out treatment with 20 nM to 100 μM PMA. The chemiluminescence responses were found to be faster for the cells treated with higher concentrations of PMA. The time scale of the oxidative response monitored using the chemiluminescence method correlated with that recorded using a microelectrode. The slow time scale of ROS production reflects the differentiation rate of THP-1 into macrophages. The effect of inhibitors on NADPH oxidase and NO synthase were also examined to find that the superoxide anion is the main radical produced from NADPH oxidase.
AB - Human monocytic leukemia cell line THP-1 differentiates into macrophages by phorbol myristate acetate (PMA) treatment. We report real-time monitoring of reactive oxygen species (ROS) production during the differentiation process. The production of ROS by THP-1 with several hours time scale has been detected using electrochemical and chemiluminescence methods. The increase in oxidation current derived from ROS arising from THP-1 was observed between 1 and 10 h after the commencement of treatment with 20 nM PMA. The response was unusually slow compared to the quick oxidative response observed in the macrophages. Chemiluminescence methods were used to further investigate the ROS production rate by carrying out treatment with 20 nM to 100 μM PMA. The chemiluminescence responses were found to be faster for the cells treated with higher concentrations of PMA. The time scale of the oxidative response monitored using the chemiluminescence method correlated with that recorded using a microelectrode. The slow time scale of ROS production reflects the differentiation rate of THP-1 into macrophages. The effect of inhibitors on NADPH oxidase and NO synthase were also examined to find that the superoxide anion is the main radical produced from NADPH oxidase.
KW - Chemiluminescence
KW - Differentiation
KW - Electrochemical method
KW - Reactive oxygen species
KW - THP-1
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U2 - 10.1016/j.aca.2005.06.034
DO - 10.1016/j.aca.2005.06.034
M3 - Article
AN - SCOPUS:23844512252
VL - 549
SP - 14
EP - 19
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
SN - 0003-2670
IS - 1-2
ER -