摘要
目的 观察腺病毒介导的LacZ基因在培养许旺细胞中的表达。 方法 用报告基因LacZ重组腺病毒感染原代培养的许旺细胞 ,X gal组织化学染色检测LacZ基因的表达。 结果 用LacZ重组腺病毒感染培养的许旺细胞时 ,有较好的量效关系和时效关系。当感染增殖率分别为 2 0 0、10 0时 ,2 4、48h后接近 10 0 %的许旺细胞被感染。而且感染后许旺细胞的形态无明显异常 ,增殖能力也没有受到影响。 结论 腺病毒介导的LacZ基因可转入体外培养的许旺细胞并高效表达 ,同时许旺细胞的生长特性并不受影响 ,为腺病毒介导神经营养因子等基因治疗促进外周周围神经损伤再生奠定了基础。
Objective To investigate the expression of LacZ gene in Schwann cells(SCs) introduced by adenoviral vector in vitro. Methods The primary culture and purification of SCs was established and the efficiency of transfection of Ad-LacZ was determined by X-gal histochemical staining in SCs. Results Ad-LacZ infected SCs in a dose-dependant and time-dependant manner. As MOI (multiplicity of infection) increased, there were increasing numbers of infected cells, so that nearly 100% of the cells were blue following X-gal staining at an MOI of 200 twenty-four hours after infection. As time period of infection prolonged, there were also increasing numbers of infected cells, so that nearly all the cells were blue at an MOI of 100 forty-eight hours after infection. The MTT test showed no significant difference among the cell groups infected with or without different MOI Ad-LacZ. Conclusions (1)adenoviral vector can transfer LacZ gene into SCs in culture efficiently; (2)the viability of infected SCs was not affected. The results suggest that neurotrophic factors could be introduced into SCs by adenoviral vector to promote peripheral nerve regeneration.
出处
《中华显微外科杂志》
CSCD
北大核心
2003年第3期198-200,共3页
Chinese Journal of Microsurgery
