Observation of Positively Charged Magnetic Nanoparticles inside HepG2 Spheroids Using Electron Microscopy

Cell Medicine

Satoshi Suzuki¹  Hisashi Iwata²  Koichi Oishi³  Yumie Koshidaka³  Yoshitaka Miyamoto³  Hiroshi Yukawa³  Shuji Hayashi³  Noritada Kaji⁴  Yoshinobu Baba⁴  Takeshi Ikeya⁵  Hirofumi Noguchi⁶  Katsutoshi Murase⁷  Hiroaki Saito⁷ 

[1] #Research Laboratories, HAB Research Organization, Ichikawa General Hospital, Chiba, Japan;**Department of Orthopaedic Surgery, Nagoya Kyoritsu Hospital, Nakagawa-ku, Nagoya, Japan;*Department of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine, Showa-ku, Nagoya, Japan;§Department of Applied Chemistry, Nagoya University Graduate School of Engineering, Nagoya University, Chikusa-ku, Nagoya, Japan;¶Photosensitive Materials Research Center, Toyo Gosei Co., Ltd., Kamimyoden, Ichikawa, Japan;†Department of Surgery, Clinical Research Center, Chiba-East Hospital, National Hospital Organization, Chiba, Japan;‡Nagoya Research Laboratory, MEITO Sangyo Co., Ltd., Nishibiwajima, Kiyosu, Japan

关键词: Magnetic iron oxide nanoparticle;    Cationic dextran;    Human hepatocellular carcinoma cells (HepG2);    Spheroids;    Electron microscopy;   

DOI  :  10.3727/215517913X666530

学科分类：生物科学（综合）

来源: Cognizant Communication Corporation

【 摘 要 】

Magnetic resonance imaging (MRI) using magnetic nanoparticles has been used to diagnose vascular diseases as well as to monitor transplanted cells and tissues. In this study, we synthesized magnetic iron oxide nanoparticles (TMADM-03), electrically charged by the presence of a cationic end-group substitution of dextran, and observed these nanoparticles inside three-dimensional models of HepG2 spheroids, which mimic tissues. Patterned cell array glass disks were prepared to visualize the presence of TMADM-03 uptaken by HepG2 spheroids using transmission electron microscopy (TEM). The HepG2 cells (2 × 105 cells) were inoculated onto Cell-able™ 12-well plates. After 48 h of culture, the cells were incubated with 75 μg Fe/ml TMADM-03 in culture medium for 24 h. To investigate the cellular function of the HepG2 spheroids, the albumin secretion was evaluated by an ELISA. The albumin secretion after incubation for 24 h was reduced compared with the secretion prior to the addition of TMADM-03. TEM image samples were prepared in a planar direction or a vertical direction to the HepG2 spheroids on patterned cell array glass disks. The incorporation of TMADM-03 inside the HepG2 spheroids was confirmed. In addition, TMADM-03 could be observed in the deeper layers of the spheroids, and this was localized in the lysosomes. These data suggest that the novel magnetic iron oxide nanoparticles invade three-dimensional HepG2 spheroids.

【 授权许可】

CC BY