Culture medium type affects endocytosis of multi-walled carbon nanotubes in BEAS-2B cells and subsequent biological response
Culture medium type affects endocytosis of multi-walled carbon nanotubes in BEAS-2B cells and subsequent biological response
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DOI：
10.1016/j.tiv.2013.04.012
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发表时间：
2013-09-01
影响因子：
3.2
通讯作者：
Kato, Hiroyuki
中科院分区：
文献类型：
--
作者：
Haniu, Hisao;Saito, Naoto;Kato, Hiroyuki
我们检查了多壁碳纳米管 (MWCNT) 的细胞毒性以及 BEAS-2B 细胞或正常人支气管上皮细胞 (HBEpCs) 在两种类型的培养基（含 10% FBS 的 Ham's F12 [Ham's F12] 和无血清生长培养基 [SFGM]）中产生的细胞因子分泌。通过荧光显微镜观察细胞对 MWCNT 的摄取，并使用流式细胞术进行分析。此外，我们评估了 MWCNT 的摄取是否被 2 种内吞作用抑制剂抑制。我们发现，在Ham's F12中培养的BEAS-2B细胞和在SFGM中培养的HBEpC表现出相似的生物学反应，但在SFGM中培养的BEAS-2B细胞没有内化MWCNT，并且50％抑制浓度值，即细胞毒性增加了10倍以上。在 Ham's F12 中培养的 BEAS-2B 细胞和 SFGM 中培养的 HBEpC 中，网格蛋白介导的内吞作用抑制剂和小窝介导的内吞作用抑制剂抑制了 MWCNT 的摄取。总之，我们建议使用含血清培养基中培养的BEAS-2B细胞作为正常人支气管上皮细胞的模型来进行纳米材料的安全性评价。然而，培养基成分可能会影响细胞质膜上表达的蛋白质，从而影响对多壁碳纳米管的生物反应。 (C) 2013 作者。由爱思唯尔有限公司出版。保留所有权利。
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. (C) 2013 The Authors. Published by Elsevier Ltd. All rights reserved.