J Korean Soc Magn Reson Med.  2012 Apr;16(1):47-54. 10.13104/jksmrm.2012.16.1.47.

In vitro MRI and Characterization of Rat Mesenchymal Stem Cells Transduced with Ferritin as MR Reporter Gene

• Shin CI
• Lee W
• Woo JS
• Park EA
• Kim PK
• Song HB
• Kim HS

• KMID: 2099837
• DOI: http://doi.org/10.13104/jksmrm.2012.16.1.47

Abstract

PURPOSE
This study was performed to evaluate the characteristics of rat mesenchymal stem cells (RMSCs) transduced with human ferritin gene and investigate in vitro MRI detectability of ferritin-transduced RMSCs.
MATERIALS AND METHODS
The RMSCs expressing both myc-tagged human ferritin heavy chain subunit (myc-FTH) and green fluorescence protein (GFP) were transduced with lentiviurs. Transduced cells were sorted by GFP expression using a fluorescence-activated cell sorter. Myc-FTH and GFP expression in transduced cells were detected by immunofluorescence staining. The cell proliferative ability and viability were assessed by MTT assay. The RMSC surface markers (CD29+/CD45-) were analyzed by flow cytometry. The intracellular iron amount was measured spectrophotometically and the presence of ferritin-iron accumulation was detected by Prussian blue staining. In vitro magnetic resonance imaging (MRI) study of cell phantoms was done on 9.4 T MR scanner to evaluate the feasibility of imaging the ferritin-transduced RMSCs.
RESULTS
The myc-FTH and GFP genes were stably transduced into RMSCs. No significant differences were observed in terms of biologic properties in transduced RMSCs compared with non-transduced RMSCs. Ferritin-transduced RMSCs exhibited increased iron accumulation ability and showed significantly lower T2 relaxation time than non-transduced RMSCs.
CONCLUSION
Ferritin gene as MR reporter gene could be used for non-invasive tracking and visualization of therapeutic mesenchymal stem cells by MRI.

Keyword

Rat mesenchymal stem cell; Reporter gene; Ferritin; Magnetic resonance imaging

MeSH Terms

Animals
Apoferritins
Ferritins
Ferrocyanides
Flow Cytometry
Fluorescence
Fluorescent Antibody Technique
Genes, Reporter
Humans
Iron
Magnetic Resonance Imaging
Mesenchymal Stromal Cells
Rats
Relaxation
Track and Field
Apoferritins
Ferritins
Ferrocyanides
Iron

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