Culture medium type affects endocytosis of multi-walled carbon nanotubes in BEAS-2B cells and subsequent biological response
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- 作者：Hisao Haniu ; Naoto Saito ; Yoshikazu Matsuda ; Tamotsu Tsukahara ; Kayo Maruyama ; Yuki Usui ; Kaoru Aoki ; Seiji Takanashi ; Shinsuke Kobayashi ; Hiroki Nomura ; Masanori Okamoto ; Masayuki Shimizu ; Hiroyuki Kato
- 关键词：AB ; Alamar blue ; CNT ; carbon nanotube ; DIC ; differential interference contrast ; FCM ; flow cytometry ; F-DPBS ; Dulbecco&rsquo ; s-PBS containing 10 % FBS ; Ham&rsquo ; s F12 ; Ham&rsquo ; s F12 containing 10 % fetal bovine serum ; H33342 ; bisbenzimide H33342 fluorochr
- 刊名：Toxicology in Vitro
- 出版年：2013
- 出版时间：September, 2013
- 年：2013
- 卷：27
- 期：6
- 页码：1679-1685
- 全文大小：1271 K
文摘
We examined the cytotoxicity of multi-walled carbon nanotubes (MWCNTs) and the resulting cytokine secretion in BEAS-2B cells or normal human bronchial epithelial cells (HBEpCs) in two types of culture media (Ham¡¯s F12 containing 10 % FBS [Ham¡¯s F12] and serum-free growth medium [SFGM]). Cellular uptake of MWCNT was observed by fluorescent microscopy and analyzed using flow cytometry. Moreover, we evaluated whether MWCNT uptake was suppressed by 2 types of endocytosis inhibitors. We found that BEAS-2B cells cultured in Ham¡¯s F12 and HBEpCs cultured in SFGM showed similar biological responses, but BEAS-2B cells cultured in SFGM did not internalize MWCNTs, and the 50 % inhibitory concentration value, i.e., the cytotoxicity, was increased by more than 10-fold. MWCNT uptake was suppressed by a clathrin-mediated endocytosis inhibitor and a caveolae-mediated endocytosis inhibitor in BEAS-2B cells cultured in Ham¡¯s F12 and HBEpCs cultured in SFGM. In conclusion, we suggest that BEAS-2B cells cultured in a medium containing serum should be used for the safety evaluation of nanomaterials as a model of normal human bronchial epithelial cells. However, the culture medium composition may affect the proteins that are expressed on the cytoplasmic membrane, which may influence the biological response to MWCNTs.