PLoS One. 2014 Nov 3;9(11):e110142.

Novel positively charged nanoparticle labeling for in vivo imaging of adipose tissue-derived stem cells.

 

Yukawa H1, Nakagawa S2, Yoshizumi Y2, Watanabe M3, Saito H4, Miyamoto Y5, Noguchi H6, Oishi K7, Ono K7, Sawada M7, Kato I4, Onoshima D8, Obayashi M1, Hayashi Y2, Kaji N9, Ishikawa T2, Hayashi S5, Baba Y10.
  • 1Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
  • 2Department of Medical Technology, Nagoya University, Graduate School of Medicine, Daikominami, Higashi-ku, Nagoya 461-8673, Japan.
  • 3Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
  • 4Nagoya Research Laboratory, MEITO Sangyo Co., Ltd., Kiyosu 452-0067, Japan.
  • 5Department of Advanced Medicine in Biotechnology and Robotics, Graduate School of Medicine, Nagoya University, Higashi-ku, Nagoya 461-0047, Japan.
  • 6Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, 207 Uehara, Nishihara, Okinawa 903-0215, Japan.
  • 7Research Institute of Environmental Medicine, Stress Adaption and Protection, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8601, Japan.
  • 8Institute of Innovative for Future Society, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
  • 9Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan; Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
  • 10Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan; Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan; Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Hayashi-cho 2217-14, Takamatsu 761-0395, Japan.

 

Abstract

Stem cell transplantation has been expected to have various applications for regenerative medicine. However, in order to detect and trace the transplanted stem cells in the body, non-invasive and widely clinically available cell imaging technologies are required. In this paper, we focused on magnetic resonance (MR) imaging technology, and investigated whether the trimethylamino dextran-coated magnetic iron oxide nanoparticle -03 (TMADM-03), which was newly developed by our group, could be used for labeling adipose tissue-derived stem cells (ASCs) as a contrast agent. No cytotoxicity was observed in ASCs transduced with less than 100 µg-Fe/mL of TMADM-03 after a one hour transduction time. The transduction efficiency of TMADM-03 into ASCs was about four-fold more efficient than that of the alkali-treated dextran-coated magnetic iron oxide nanoparticle (ATDM), which is a major component of commercially available contrast agents such as ferucarbotran (Resovist), and the level of labeling was maintained for at least two weeks. In addition, the differentiation ability of ASCs labeled with TMADM-03 and their ability to produce cytokines such as hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF) and prostaglandin E2 (PGE2), were confirmed to be maintained. The ASCs labeled with TMADM-03 were transplanted into the left kidney capsule of a mouse. The labeled ASCs could be imaged with good contrast using a 1T MR imaging system. These data suggest that TMADM-03 can therefore be utilized as a contrast agent for the MR imaging of stem cells.

PMID: 25365191

 

Supplemental documents

fig1

Figure 1. A: Schematic diagrams (left) and the chemical formulas (right) of ATDM (a) and TMADM-03 (b). B: SEM images of ATDM (a) and TMADM-03.

We recently reported the development of six kinds of trimethylamino dextran-coated magnetic iron oxide nanoparticles with different positive charges (TMADM-01~05). The characteristics of these particles, such as their diameter, zeta voltage, and cationic end-group substitution degree, have all been previously measured and compared [1]. In this paper, the chemical formula (Fig. 1A), SEM image (Fig. 1B) of ATDM and TMADM-03 were compared. The SEM images and particle side distributions of ATDM and TMADM-03 showed that they had almost the similar circle shape and size. As a result, the labeling efficiency of TMADM-03 for ASCs could be confirmed to be higher than that of ATDM, and was effective at least for two weeks.

fig2

Figure 2. TMADM-03 particles were mainly transduced into ASCs by the macropinocytosis pathway.

 

 

To verify the uptake mechanism of TMADM-03 in ASCs, the cells were treated with endocytosis inhibitors such as sodium azide and 2-deoxy-D-glucose (endocytosis inhibitor), amiloride (an inhibitor of Na+/H+ exchanger required for macropinocytosis), filipin III (an inhibitor of caveolae formation by binding with cholesterol), or chlorpromazine (CPZ: an inhibitor of AP-2 mediated clathrin-coated pit formation) at 37°C for 1 h (15 min for amiloride). In addition, the treatment of incubation at 4°C for 1 h was conducted for endocytosis inhibition. The transduction of TMADM-03 was inhibited by the incubation at 4°C and the treatments of sodium azide and 2-deoxy-D-glucose and amiloride. These data suggest that the uptake of TMADM-03 in ASCs was mainly dependent on the macropinocytosis in endocytosis (Fig. 2).

Finally, a schematic representation of this study of the labeling and in vivo MR imaging of stem cells using positively charged TMADM-03 was shown (Fig.3).

 

fig3

Figure 3. A schematic representation of this study of the labeling and in vivo MR imaging of stem cells using positively charged TMADM-03.

 

 

Reference:

  1. Oishi, K.; Noguchi, H.; Saito, H.; Yukawa, H.; Miyamoto, Y.; Ono, K.; Murase, K.; Sawada, M.; Hayashi, S. Novel Positive-charged Nanoparticles for Efficient Magnetic Resonance Imaging of Islet Transplantation. Cell medicine 2012, 3, 43-49.
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