Superparamagnetic Iron Oxide Labeling of Spinal Cord Neural Stem Cells Genetically Modified by Nerve Growth Factor-β

• Published in: Journal of Huazhong University of Science and Technology. Medical sciences 2009-04, Vol.29 (2), p.235-238
• Main Author: 雷德强 赵洪洋 邓兴力 刘如恩 张方成 姚东晓
• Format: Article
• Language: English
• Subjects: Animals; blotting; Cells, Cultured; Dextrans; Embryo, Mammalian; Magnetic Resonance Imaging; Magnetics; Magnetite Nanoparticles; Medicine; Medicine & Public Health; Nerve Growth Factor - genetics; Nerve Growth Factor - pharmacology; Neural Stem Cells - cytology; Rats; Spinal Cord - cytology; Transfection; 免疫细胞化学; 基因修饰; 神经生长因子; 脊髓源性神经干细胞; 脊髓神经干细胞; 超顺磁性氧化铁粒子; 透射电子显微镜
• ISSN: 1672-0733; 1993-1352
• DOI: 10.1007/s11596-009-0220-3

This study established superparamagnetic iron oxide （SPIO）-labeled nerve growth fac-tor-β （NGF-β） gene-modified spinal cord-derived neural stem cells （NSCs）. The El4 rat embryonic spinal cord-derived NSCs were isolated and cultured. The cells of the third passage were transfected with plasmid pcDNA3-hNGFβ by using FuGENE HD transfection reagent. The expression of NGFβ was measured by immunocytochemistry and Western blotting. The positive clones were selected, allowed to proliferate and then labeled with SPIO, which was mediated by FuGENE HD transfection reagent. Prussian blue staining and transmission electron microscopy （TEM） were used to identify the SPIO particles in the cells. The distinctive markers for stem cells （nestin）, neuron （β-Ⅲ-tubulin）, oligodendrocyte （CNPase） and astrocyte （GFAP） were employed to evaluate the differentiation ability of the labeled cells. The immunocytochemistry and western blotting showed that NGF-β was expressed in spinal cord-derived NSCs. Prussian blue staining indicated that numerous blue-stained particles appeared in the cytoplasma of the labeled cells. TEM showed that SPIO particles were found in vacuolar structures of different sizes and the cytoplasma. The immunocytochemistry demonstrated that the labeled cells were nestin-positive. After differentiation, the cells expressed β-Ⅲ-tubulin, CNPase and GFAP. It was concluded that the SPIO-labeled NGF-β gene-modified spinal cord-derived NSC were successfully established, which are multipotent and capable of self-renewal.