In vitro MR imaging of Fe2O3-arginine labeled heNOS gene modified endothelial progenitor cells
10.3321/j.issn:0253-3758.2008.08.006
- VernacularTitle:磁探针标记人内皮型一氧化氮合酶基因修饰的内皮祖细胞及其体外磁共振成像实验研究
- Author:
Fang NIE
1
;
Xiao-Li MAI
;
Jun CHEN
;
Ning GU
;
Hong-Jian SHI
;
Ai-Hong CAO
;
Yu-Qing GE
;
Yu ZHANG
;
Gao-Jun TENG
Author Information
1. 东南大学附属中大医院
- Keywords:
Endothelial progenitor cells;
Gene transfer;
Magnetic resonance imaging;
Labeling
- From:
Chinese Journal of Cardiology
2008;36(8):695-701
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the feasibility of in vitro magnetic resonance imaging on Fe2O3-arginine labeled heNOS gene modified endothelial progenitor cells(EPCs). Methods Fe2O3 was incubated with arginine to form Fe2O3-arginine complex. Rabbit peripheral blood mononuclear cells (MNCs)were isolated and EPCs were isolated by adherence method, expanded and modified with heNOS gene using Lipofectamine<'TM2000. After 48 hours, genetically modified EPCs were incubated with Fe2O3-arginine for 24 hours. Intracellular iron was detected by Prussian blue stain. The expression of heNOS gene was detected by Western blot. MTT assay was used to evaluate cell survival and proliferation of Fe2O3-arginine labeled heNOS-EPCs. Flow cytometry was used to measure cell apoptosis. The cells underwent in vitro MR imaging with various sequences. Results Iron-containing intracytoplasmatic vesicles could be clearly observed with Prussian blue staining, and the labeling rate of labeled heNOS-EPCs were similar to that of labeled EPCs (around 100% ). Survival and apoptosis rates obtained by MTT and flow cytometry analysis were similar among labeled heNOS-EPCs, labeled EPCs and unlabeled EPCs with Fe2O3-arginine. The signal intensity on MRI was equally decreased in labeled heNOS-EPCs and labeled EPCs compared with that in unlabeled cells. The percentage change in signal intensity (ASI) was most significant on T2 * WI and △SI was significantly lower in cells labeled for 7 days than that labeled for 1 days. Conclusions The beNOS gene can be successfully transfected into rabbit peripheral blood EPCs using LipofectamineTM2000. The heNOS-EPCs can be labeled with Fe2O3-arginine without significant change in viability and proliferation capacity. The labeled heNOS-EPCs can be imaged with standard 1.5 T MR equipment. The degree of MR signal intensity may indirectly reflect the cell count, growth and division status.
