You are here
Dual transfer of GFP gene and MGd into stem-progenitor cells: toward in vivo MRI of stem cell-mediated gene therapy of atherosclerosis.
Title | Dual transfer of GFP gene and MGd into stem-progenitor cells: toward in vivo MRI of stem cell-mediated gene therapy of atherosclerosis. |
Publication Type | Journal Article |
Year of Publication | 2010 |
Authors | Qiu, B, Treuting, P, Zhan, X, Xie, D, Frevert, CW, Yang, X |
Journal | Acad Radiol |
Volume | 17 |
Issue | 5 |
Pagination | 547-52 |
Date Published | 2010 May |
ISSN | 1878-4046 |
Keywords | Animals, Atherosclerosis, Cell Line, Contrast Media, Green Fluorescent Proteins, Magnetic Resonance Imaging, Mesenchymal Stem Cell Transplantation, Metalloporphyrins, Mice, Mice, Transgenic, Surgery, Computer-Assisted |
Abstract | RATIONALE AND OBJECTIVES: The aim of this study was to develop a new technique, the use of magnetic resonance (MR) imaging (MRI) to monitor gene/MR-cotransferred stem-progenitor cells (SPCs) recruited to atherosclerosis. MATERIALS AND METHODS: First, a green fluorescent protein (GFP) gene and a T1 MR contrast agent (motexafin gadolinium [MGd]) were cotransferred into neural or bone marrow (BM)-derived SPCs. GFP expression and MGd signal were confirmed by fluorescent microscopy and quantified by flow cytometry. Cell viability and proliferation were then evaluated by trypan blue exclusion and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay, and GFP/MGd-transferred cells were imaged using 1.5-T and 9.4-T MR scanners. For in vivo validation, GFP/MGd-cotransferred beta-galactosidase-BM SPCs were transplanted to apolipoprotein E-knockout mice, and cell migration to atherosclerotic aortas was monitored using 9.4-T micro-MRI with subsequent histologic correlations. RESULTS: Fluorescent microscopy demonstrated simultaneous GFP expression and MGd signals in cotransferred-cells. Quantitative flow cytometry showed GFP-positive cells at 47 +/- 25% and 56 +/- 12% and MGd-positive cells at 96 +/- 6% and 57 +/- 11% for neural stem cells and BM cells, respectively. Cell viability and metabolic rates of cotransferred cells were 86 +/- 4% and 84 +/- 12%, respectively. In vivo MRI revealed high MR signals of the aortic walls in GFP/MGd-transferred mice, which were confirmed by histologic correlations. CONCLUSION: This study has initially proven the new concept of MRI for plaque-specific, cell-mediated gene expression of atherosclerosis. |
DOI | 10.1016/j.acra.2010.02.004 |
Alternate Journal | Acad Radiol |
PubMed ID | 20227305 |
Grant List | R01 HL078672 / HL / NHLBI NIH HHS / United States |