Molecular Imaging and Biology

, Volume 13, Issue 3, pp 443–451

Direct Labeling of hMSC with SPIO: the Long-Term Influence on Toxicity, Chondrogenic Differentiation Capacity, and Intracellular Distribution

  • Chung-Yi Yang
  • Jong-Kai Hsiao
  • Ming-Fong Tai
  • Shin-Tai Chen
  • Hui-Ying Cheng
  • Jaw-Lin Wang
  • Hon-Man Liu
Research Article

DOI: 10.1007/s11307-010-0360-7

Cite this article as:
Yang, CY., Hsiao, JK., Tai, MF. et al. Mol Imaging Biol (2011) 13: 443. doi:10.1007/s11307-010-0360-7

Abstract

Purpose

The purpose of this study was to evaluate the long-term cellular toxicity, labeling efficiency, chondrogenic differentiation capacity, and intracellular distribution following direct superparamagnetic iron oxide (SPIO) nanoparticle labeling of human mesenchymal stem cells (hMSCs) in the absence of transfection agents.

Procedures

hMSCs were incubated with a SPIO, Ferucarbotran, at concentrations of 0, 1, 10, and 100 μg Fe/ml for 24 or 72 h. The cell granularity and size change, reactive oxygen species generation, and mitochondria membrane potential were measured by flow cytometry. The differentiation capacity of the cells into chondrocytes was determined by Alcian blue and Safranin-O staining, immunocytochemical analysis, and reverse transcription polymerase chain reaction.

Results

The intracellular distribution of the internalized particles was visualized via confocal microscopy. No significant difference was found in the toxicity of labeled cells relative to controls. Successful chondrogenesis of Ferucarbotran-labeled hMSCs was confirmed. The intracellular SPIO nanoparticles were located within the lysosomes.

Conclusions

In conclusion, we have demonstrated the feasibility of direct labeling with Ferucarbotran without impairment of cellular function, toxicity, or inhibition of differentiation capacity. Furthermore, lysosomal metabolism takes place after intracellular uptake of Ferucarbotran.

Key Words

Ferucarbotran Mesenchymal stem cells Chondrogenesis 

Abbreviations

hMSC

human mesenchymal stem cell

MSC

mesenchymal stem cell

PLL

poly-l-lysine

SPIO

superparamagnetic iron oxide

hTERT

human telomerase reverse transcriptase

DMEM

Dulbecco’s modified Eagle’s medium

FBS

fetal bovine serum

ROS

reactive oxygen species

DCFDA

dichlorofluoscein diacetate

MMP

mitochondria membrane potential

PBS

phosphate-buffered saline

Copyright information

© Academy of Molecular Imaging and Society for Molecular Imaging 2010

Authors and Affiliations

  • Chung-Yi Yang
    • 1
    • 2
    • 3
  • Jong-Kai Hsiao
    • 3
  • Ming-Fong Tai
    • 4
  • Shin-Tai Chen
    • 5
  • Hui-Ying Cheng
    • 3
  • Jaw-Lin Wang
    • 1
  • Hon-Man Liu
    • 3
    • 6
  1. 1.Institute of Biomedical EngineeringNational Taiwan UniversityTaipeiTaiwan
  2. 2.Department of Medical ImagingNational Taiwan University Hospital Yun-Lin BranchDouliuTaiwan
  3. 3.Department of Medical ImagingNational Taiwan University Hospital and National Taiwan University College of MedicineTaipeiTaiwan
  4. 4.Department of PhysicsNational Tsing Hua UniversityHsinchuTaiwan
  5. 5.Department of Biochemistry, Musculoskeletal Disease Center, J.L. Pettis VA Medical CenterLoma Linda UniversityLoma LindaUSA
  6. 6.Department of RadiologyNational Taiwan University Hospital and College of MedicineTaipeiTaiwan

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