Construction of 3-D cellular multi-layers with extracellular matrix assembly using magnetic nanoparticles

Yu Bin Lee, Joong Yup Lee, Taufiq Ahmad, Seongwoo Bak, Jinkyu Lee, Hwan D. Kim, Tai Hyun Park, Nathaniel S. Hwang, Youn Mook Lim, Heungsoo Shin

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Construction of 3-dimensional (3-D) engineered tissue is increasingly being investigated for use in drug discovery and regenerative medicine. Here, we developed multi-layered 3-D cellular assembly by using magnetic nanoparticles (MNP) isolated from Magnetospirillum sp. AMB-1 magnetotactic bacteria. Magnetized human dermal fibroblasts (HDFBs) were prepared by treatment with the MNP, induced to form 3-D assembly under a magnetic field. Analyses including LIVE/DEAD assay, transmission electron microscopy revealed that the MNP were internalized via clathrin-mediated endocytosis without cytotoxicity. The magnetized HDFBs could build 3-D structure as a function of seeding density. When the highest seeding density (5×105 cells/mm2) was used, the thickness of assembly was 41.90±1.69μm, with approximately 9.3±1.6 cell layers being formed. Immunofluorescence staining confirmed homogeneous distribution of ECM and junction proteins throughout the 3-D assembly. Real-time PCR analysis showed decrease in expression levels of collagen types I and IV but increase in that of connexin 43 in the 3-D assembly compared with the 2-D culture. Finally, we demonstrated that the discernible layers can be formed hierarchically by serial assembly. In conclusion, our study showed that a multi-layered structure can be easily prepared using magnetically-assisted cellular assembly with highlighting cell-cell and cell-ECM communication.

Original languageEnglish
Pages (from-to)1916-1928
Number of pages13
JournalJournal of Biomedical Nanotechnology
Volume12
Issue number10
DOIs
StatePublished - Oct 2016

Bibliographical note

Funding Information:
This work was supported by a National Research Foundation of Korea (NRF) funded by the the Ministry of Science, ICT and Future Planning (NRF-2013R1A2A2A03067809) and Radiation Technology R&D program (NRF-2015M2A2A6021052).

Publisher Copyright:
Copyright © 2016 American Scientific Publishers All rights reserved.

Keywords

  • 3-D Cellular Assembly
  • Cellular Internalization
  • Hierarchical Tissue Mimicry
  • Magnetic Nanoparticles
  • Multi-Layered Structure

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