摘要
目的观察阳离子脂质体转染神经营养因子-3(NT-3)基因在培养雪旺细胞(Schwann cell,SC)的表达和提高雪旺细胞分泌NT-3的能力。方法用消化后的组织块法和差速离心法分离、纯化雪旺细胞。用脂质体转染法将NT-3基因导人雪旺细胞,ELISA双抗体夹心法检测NT-3的表达;免疫组化S-100法鉴定SC的纯度。结果经免疫组化S-100法鉴定SC的纯度达95%以上。ELISA双抗体夹心法测定转染后不同时期的NT-3的浓度,2周后表达增加,4周后明显增加,差异有统计学意义(P<0.01)。结论NT-3基因可转入培养的SC并有高效表达。SC作为受体细胞,易于获取并能在体外大量培养繁殖;能较长时间表达所携带基因而不衰减,并能稳定地大量表达。
Objective The purpose of this study is to observe NT-3 expression in cultured Schwann cells （SCs） that are genetically modified by human NT-3 genes, and to increase secretion of NT-3 in those cells. Methods SCs were separated and purified with post-digestion tissue blot and time-spced difference stick. Plasmids expressing NT-3 genes were then transfected to those SCs with lipofectamine. At different phases of gene transfection, NT-3 expression levels in conditioned medium of SCs were measured with enzyme-linked immunoassay sensitive assay （ELISA） and the purity of SCs determined by S-100 immunohistochemistry. Results The purity Of SCs was 95% by S-100 immunohistochemistry. Compared with non-transduced SCs, the level of NT-3 in transfeeted SCs began to increase after two weeks of gene transfer, and increased obviously after four weeks. The results had statistical significance （ P 〈 0.01 ）. Conclusion NT-3 gene can be effectively transduced into cultured SCs. As receptor cells, SCs can be easily obtained and cultured and massively reproduced in vitro. They can be transduced to secrete augmented levels of NT-3 and express the transgene for a prolonged period.
出处
《中华手外科杂志》
CSCD
北大核心
2007年第2期107-110,共4页
Chinese Journal of Hand Surgery
