摘要
目的探讨人脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)基因工程细胞的建立及其对体外培养的小鼠螺旋神经元生长活性的影响。方法参照分子克隆技术成功构建BDNF真核表达载体———pcD-NA3.1(-)-BDNF,体外证实BDNF基因转染猿猴病毒40肝脏转化细胞(COS7)后正常表达,将骨髓基质细胞分离培养,体外转染骨髓基质细胞建立BDNF基因工程细胞,观察该基因工程细胞对体外培养小鼠螺旋神经元的生长活性的影响。结果成功构建BDNF基因真核表达载体,并且建立了BDNF基因工程细胞;在体外BDNF基因工程细胞能促进螺旋神经元的生长,并保护螺旋神经元免受氧化损伤。结论建立的BDNF基因修饰的骨髓基质细胞,对体外螺旋神经元生长、生存起着重要的保护作用,为进一步研究基因修饰的骨髓基质细胞内耳移植奠定了重要的基础。
Ojective To establish human brain - derived neurotrophic factor （BDNF） genetical modified marrow stremal cells and to evaluate the influence on the viability of the cultured spiral ganglion cells from Kunmirlg mice. Methods Eukaryotic expression plasmid pcDNA3.1 （ - ） - BDNF was constructed according to the molecular clone approach, Marrow stremal cells （MSC） were separated and cultured. BDNF genetical modified marrow stromal cells were established and verified by biochemical analysis. The bio- effect to spiral ganglion cells, especially with the oxidative damage under the different concentration of H2O2 was observed. Results PcDNA3.1 （ - ） - BDNF and BDNF genetical engineering cells were successfully established. The supernatant from BDNF - MSCs remarkably improved the survival rate of the spiral ganglion cells. Conclusion BDNF genetical modified marrow stromal cells are successfully established, and the genetical engineering cell plays an important role in the protection the spiral ganglion cells from the oxidative damage. This study provide a strong basis for genetical engineering cells transplantation into the inner ear.
出处
《听力学及言语疾病杂志》
CAS
CSCD
2006年第2期121-121,122-124,T0001,共5页
Journal of Audiology and Speech Pathology
基金
国家自然科学基金资助(编号30070807)
关键词
基因工程细胞
骨髓基质干细胞
脑源性神经营养因子
螺旋神经元
Genetical engineering cells
Marrow stromal cell
Brain- derived neurotrophic factor（BDNF）
Spiral ganglion cell
