Construction of Multi-layered Cell Sheet Using Magnetite Nanoparticles and Magnetic Force

  • Akira Ito
  • Hiroyuki Honda
  • Masamichi Kamihira
Conference paper
Part of the Animal Cell Technology: Basic & Applied Aspects book series (ANICELLTECH, volume 15)


Novel technologies to establish 3D tissue-like constructs are desired for tissue engineering. In the present study, magnetic force and magnetite nanoparticles were used to construct a layered mesenchymal stem cell (MSC) sheet, a layered cardiomyocyte sheet, and a layered fibroblast sheet involving capillaries. Magnetite cationic liposomes (MCLs), which have a positive surface charge, were taken up by the target cells. When a magnet was set under a tissue culture dish, magnetically labeled target cells were attracted and then adhered to form a layered cell sheet. For MSC sheets, MSCs within the sheets maintained an in vitro multi-differentiation ability into osteoblasts, adipocytes or chondrocytes after a 21-day culture period using each induction medium. By using an electromagnet, MSC sheets were harvested and transplanted into the bone defect in cranium of nude rats. Histological observation revealed that new bones surrounded by osteoblast-like cells were formed in the defect area of rats on 14 days after transplantation with MSC sheets. For cardiomyocyte sheets, the immuno-fluorescence staining of connexin43 revealed the existence of gap junctions within the cardiomyocyte sheets. Moreover, electrical connection within the cardiomyocyte sheets was confirmed by using extracellular potential mapping. For fibroblast sheets, normal human dermal fibroblasts (NHDFs) sheets contained the major dermal extracellular matrix (ECM) components (fibronectin and type I collagen). Human umbilical vein endothelial cells (HUVECs) were co-cultured with NHDF sheets, resulted in tube-like formation of HUVECs, resembling early capillaries, within NHDF sheets after short-term 3D co-culture. These results suggest that this novel use of magnetite nanoparticles and magnetic force, which we refer to as “magnetic force-based tissue engineering (Mag-TE)”, offers a major advancement in tissue engineering.


Tissue engineering magnetite nanoparticle liposome 


  1. 1.
    A. Ito, M. Hayashida, H. Honda, K. Hata, H. Kagami, M. Ueda, T. Kobayashi, Construction and harvest of multilayered keratinocyte sheets using magnetite nanoparticles and magnetic force. Tissue Eng 10(5-6), 873–880 (2004).CrossRefPubMedGoogle Scholar
  2. 2.
    A. Ito, Y. Takizawa, H. Honda, K. Hata, H. Kagami, M. Ueda, T. Kobayashi, Tissue engineering using magnetite nanoparticles and magnetic force: heterotypic layers of cocultured hepatocytes and endothelial cells. Tissue Eng 10(5-6), 833–840 (2004).CrossRefPubMedGoogle Scholar
  3. 3.
    A. Ito, E. Hibino, C. Kobayashi, H. Terasaki, H. Kagami, M. Ueda, T. Kobayashi, H. Honda, Construction and delivery of tissue-engineered human retinal pigment epithelial cell sheets, using magnetite nanoparticles and magnetic force. Tissue Eng11(3-4), 489–496 (2005).CrossRefPubMedGoogle Scholar
  4. 4.
    A. Ito, K. Ino, M. Hayashida, T. Kobayashi, H. Matsunuma, H. Kagami, M. Ueda, H. Honda, Novel methodology for fabrication of tissue-engineered tubular constructs using magnetite nanoparticles and magnetic force. Tissue Eng 11(9-10), 1553–1561 (2005).CrossRefPubMedGoogle Scholar
  5. 5.
    K. Shimizu, A. Ito, T. Yoshida, Y. Yamada, M. Ueda, H. Honda, Bone tissue engineering with human mesenchymal stem cell sheets constructed using magnetite nanoparticles and magnetic force. J Biomed Mater Res B Appl Biomater 82(2), 471–480 (2007).PubMedGoogle Scholar
  6. 6.
    K. Shimizu, A. Ito, J.K. Lee, T. Yoshida, K. Miwa, H. Ishiguro, Y. Numaguchi, T. Murohara, I. Kodama, H. Honda, Construction of multi-layered cardiomyocyte sheets using magnetite nanoparticles and magnetic force. Biotechnol Bioeng 96(4), 803–809 (2007).CrossRefPubMedGoogle Scholar
  7. 7.
    K. Ino, A. Ito, H. Kumazawa, H. Kagami, M. Ueda, H. Honda, Incorporation of capillary-like structures into dermal cell sheets constructed by magnetic force-based tissue engineering. J Chem Eng Jpn 40(1), 51–58 (2007).CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Akira Ito
    • 1
  • Hiroyuki Honda
    • 2
  • Masamichi Kamihira
    • 1
  1. 1.Department of Chemical Engineering, Faculty of EngineeringKyushu UniversityFukuokaJapan
  2. 2.Department of Biotechnology, School of EngineeringNagoya UniversityFuro-cho, Chikusa-ku, NagoyaJapan

Personalised recommendations