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Shin, Lee, Woo, Park, Kim, Song, and Kim: In vitro MRI and Characterization of Rat Mesenchymal Stem Cells Transduced with Ferritin as MR Reporter Gene
Original Article
Journal of the Korean Society of Magnetic Resonance in Medicine

Journal of the Korean Society of Magnetic Resonance in Medicine 2012; 16(1): 47-54.

Published online: 30 April 2012

DOI: https://doi.org/10.0000/jksmrm.2012.16.1.47

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

Cheong-il Shin1, Whal Lee1, 2, Ji Su Woo1, Eun-Ah Park1, 2, Pan-ki Kim2, Hyun Bok Song2, Hoe Suk Kim1, 3

1Department of Radiology, Seoul National University Hospital, Korea.

2SNU-Duke Cardiovascular MR Research Center, Seoul National University, Korea.

3Institute of Radiation Medicine, SNUMRC (Seoul National University Medical Research Center), Korea.

Corresponding author: Whal Lee, M.D. and Hoe Suk Kim, Ph.D., Department of Radiology, Seoul National University Hospital, 101 Daehangno, Jongno-gu, Seoul 110-744, Korea. Fax. 82-2-743-6385, Tel. 82-2-2072-0898, whal.lee@gmail.com for Whal Lee, M.D., Tel. 82-2-2072-1791, hoeskim@snu.ac.kr for Hoe Suk Kim, Ph.D.

Received 1 April 2012       Revised 25 April 2012       Accepted 25 April 2012

Copyright © 2012 Korean Society of Magnetic Resonance in Medicine

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.

Keywords: Rat mesenchymal stem cell, Reporter gene, Ferritin, Magnetic resonance imaging

References

1. Cohen B, Ziv K, Plaks V, et al. MRI detection of transcriptional regulation of gene expression in transgenic mice. Nature Medicine. 2007. 13:498–503.
2. Kim HS, Cho HR, Choi SH, Woo JS, Moon WK. In vivo imaging of tumor transduced with bimodal lentiviral vector encoding human ferritin and green fluorescent protein on a 1.5T clinical magnetic resonance scanner. Cancer Research. 2010. 70:7315–7324.
3. Genove G, DeMarco U, Xu H, Goins WF, Ahrens ET. A new transgene reporter for in vivo magnetic resonance imaging. Nature Medicine. 2005. 11:450–454.
4. Aung W, Hasegawa S, Koshikawa-Yano M, et al. Visualization of in vivo electroporation-mediated transgene expression in experimental tumors by optical and magnetic resonance imaging. Gene Therapy. 2009. 16:830–839.
5. Cohen B, Dafni H, Meir G, Harmelin A, Neeman M. Ferritin as an endogenous MRI reporter for noninvasive imaging of gene expression in C6 glioma tumors. Neoplasia. 2005. 7:109–117.
6. Liu J, Cheng EC, Long RC, et al. Noninvasive monitoring of embryonic stem cells in vivo with MRI transgene reporter. Tissue engineering Part C, Methods. 2009. 15:739–747.
7. Kim YJ, Huh Y-M, Choe KO, et al. In vivo magnetic resonance imaging of injected mesenchymal stem cells in rat myocardial infarction; simultaneous cell tracking and left ventricular function measurement. International Journal of Cardiovascular Imaging. 2009. 25:99–109.
8. Agudelo CA, Tachibana Y, Noboru T, Iida H, Yamaoka T. Long-term in vivo magnetic resonance imaging tracking of endothelial progenitor cells transplanted in rat ischemic limbs and their angiogenic potential. Tissue engineering Part A. 2011. 17:2079–2089.
9. Agudelo CA, Tachibana Y, Hurtado AF, Ose T, Iida H, Yamaoka T. The use of magnetic resonance cell tracking to monitor endothelial progenitor cells in a rat hindlimb ischemic model. Biomaterials. 2012. 33:2439–2448.
10. Campan M, Lionetti V, Aquaro GD, et al. Ferritin as a reporter gene for in vivo tracking of stem cells by 1.5-T cardiac MRI in a rat model of myocardial infarction. Am J Physiol Heart Circ Physiol. 2011. 300:H2238–H2250.
11. Li Y, Zhang D, Zhang Y, He G, Zhang F. Augmentation of neovascularization in murine hindlimb ischemia by combined therapy with simvastatin and bone marrow-derived mesenchymal stem cells transplantation. J Biomed Sci. 2010. 17:75.
12. Laurila JP, Laatikainen L, Castellone MD, et al. Human embryonic stem cell-derived mesenchymal stromal cell transplantation in a rat hind limb injury model. Cytotherapy. 2009. 11:726–737.
13. Iwase T, Nagaya N, Fujii T, et al. Comparison of angiogenic potency between mesenchymal stem cells and mononuclear cells in a rat model of hindlimb ischemia. Cardiovasc Res. 2005. 66:543–551.
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