In vitro MRI and Characterization of Rat Mesenchymal Stem Cells Transduced with Ferritin as MR Reporter Gene.
10.13104/jksmrm.2012.16.1.47
- Author:
Cheong Il SHIN
1
;
Whal LEE
;
Ji Su WOO
;
Eun Ah PARK
;
Pan Ki KIM
;
Hyun Bok SONG
;
Hoe Suk KIM
Author Information
1. Department of Radiology, Seoul National University Hospital, Korea. whal.lee@gmail.com, hoeskim@snu.ac.kr
- Publication Type:In Vitro ; Original Article
- Keywords:
Rat mesenchymal stem cell;
Reporter gene;
Ferritin;
Magnetic resonance imaging
- MeSH:
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
- From:Journal of the Korean Society of Magnetic Resonance in Medicine
2012;16(1):47-54
- CountryRepublic of Korea
- Language:English
-
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.
