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作 者:姚谦明[1] 徐如祥[1] 姜晓丹[1] 蔡颖谦[1]
机构地区:[1]南方医科大学珠江医院神经外科,广州在读博士研究生510282
出 处:《中华神经医学杂志》2005年第12期1194-1196,1203,共4页Chinese Journal of Neuromedicine
基 金:国家自然科学基金(30270491);广东省重大科技计划项目[粤科基办(2000)25;(2004)08;粤财企(2001)367;(2003)209;粤科技字(2004)112];军队重点科技项目(01Z054)
摘 要:目的获得表达孤儿核受体(Nurr1)基因的骨髓源性神经干细胞(BMSCs-NSCs)。方法构建携带Nurr1基因的重组腺相关病毒(AAV)载体AAV-pcDNA3.1-Nurr1,提取质粒,用脂质体转染法转染大鼠BMSCs-NSCs,并用RT-PCR和免疫细胞化学方法检测阳性细胞。结果经酶切鉴定和DNA测序,证实得到了序列正确的重组pAAV-Nurr1,获得了Nurr1阳性的BMSCs-NSCs。结论重组AAV携带的Nurr1基因能够在BMSCs-NSCs中表达,本研究为进一步探讨将该基因工程细胞用于帕金森病基因治疗提供了可能性。Objective To obtain the genetically modified bone marrow-derived neural stem cells expressing Nurr1 gene. Methods A recombinant adeno-associated virus vector encoding Nurr1 (pAAV-Nurr1) was constructed with the technique of molecular cloning and transfected into the neural stem cells derived from bone marrow of rats with Lipofectamin 2000. The positive cells were identified by the methods of immunocytochemistry and RT-PCR. Results pcDNA3, 1-Nurr1 was identified by restriction endonuclease digestion and DNA sequencing. Nurr1-positive neural stem cells infected with AAV-pcDNA 3.1-Nurr1 vector were obtained successfully. Conclusion Nurr1 gene in recombinant adeno-associated virus vector can be expressed in neural stem cells derived from bone marrow, which offers a help for the further research on gene therapy of Parkinson' s disease.
关 键 词:孤儿核受体基岗 腺相关病毒 骨髓源性神经干细胞 帕金森病
分 类 号:R329[医药卫生—人体解剖和组织胚胎学]
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