期刊
CONTRAST MEDIA & MOLECULAR IMAGING
卷 11, 期 3, 页码 236-244出版社
WILEY-HINDAWI
DOI: 10.1002/cmmi.1686
关键词
MRI; reporter gene; cell tracking; transferrin receptor; ferritin; chick embryo; CHO-K1
资金
- Marie Curie EU Grant [PERG08-GA-2010-276909]
- UK EPSRC Grant [EP/H046143]
- UK Regenerative Medicine Platform [MR/K026739/1]
- Medical Research Council (MRC) [MR/L012707/1]
- UK BBSRC [BB/F011350/1]
- Biotechnology and Biological Sciences Research Council [BB/F011350/1] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [EP/H046143/1] Funding Source: researchfish
- Medical Research Council [MR/M026256/1, MR/K026739/1, MR/L012707/1] Funding Source: researchfish
- BBSRC [BB/F011350/1] Funding Source: UKRI
- EPSRC [EP/H046143/1] Funding Source: UKRI
- MRC [MR/L012707/1, MR/K026739/1, MR/M026256/1] Funding Source: UKRI
Magnetic resonance (MR) reporter genes have the potential for tracking the biodistribution and fate of cells in vivo, thus allowing the safety, efficacy and mechanisms of action of cell-based therapies to be comprehensively assessed. In this study, we evaluate the effectiveness of the iron importer transferrin receptor-1 (TfR1) as an MR reporter gene in the model cell line CHO-K1. Overexpression of the TfR1 transgene led to a reduction in the levels of endogenous TfR1 mRNA, but to a 60-fold increase in total TfR1 protein levels. Although the mRNA levels of ferritin heavy chain-1 (Fth1) did not change, Fth1 protein levels increased 13-fold. The concentration of intracellular iron increased significantly, even when cells were cultured in medium that was not supplemented with iron and the amount of iron in the extracellular environment was thus at physiological levels. However, we found that, by supplementing the cell culture medium with ferric citrate, a comparable degree of iron uptake and MR contrast could be achieved in control cells that did not express the TfR1 transgene. Sufficient MR contrast to enable the cells to be detected in vivo following their administration into the midbrain of chick embryos was obtained irrespective of the reporter gene. We conclude that TfR1 is not an effective reporter and that, to track the biodistribution of cells with MR imaging in the short term, it is sufficient to simply culture cells in the presence of ferric citrate. Copyright (c) 2016 The Authors Contrast Media & Molecular Imaging Published by John Wiley & Sons Ltd.
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