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Study on neural stem cells differentiated into cholinergic neuron induced by TrkA gene transfection

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Author:
No author available
Journal Title:
CHINESE JOURNAL OF NEUROMEDICINE
Issue:
1
DOI:
10.3760/cma.j.issn.1671-8925.2007.01.002
Key Word:
TrkA基因;神经干细胞;分化;胆碱能神经元

Abstract: 目的 研究外源性TrkA基因修饰C17.2神经干细胞,对神经干细胞定向分化的影响. 方法 采用脂质体法将携带有TrkA基因的真核表达载体pcDNA3.1(+)/TrkA转染C17.2神经干细胞,Western blot观察转染后基因表达情况;将正常生长的C17.2神经干细胞随机分成两组(A组和C组),将重组质粒转染阳性的C17.2神经干细胞随机分成两组(B组和D组),并用100ng/mL神经生长因子(NGF)分别作用于C组及D组,应用间接免疫荧光染色方法,观察各组细胞胆碱乙酰转移酶(ChAT)的表达. 结果 Western blot结果显示转染组TrkA蛋白表达明显高于非转染组,说明外源性TrkA基因导入靶细胞,并实现蛋白表达;间接免疫荧光染色显示,经NGF孵育的转染组细胞(D组)约有26%呈ChAT阳性,而非转染组(C组)约为9%,未经NGF孵育的A、B组未观察到ChAT阳性细胞. 结论 应用外源性TrkA基因修饰神经干细胞,造成TrkA基因过度表达,在NGF作用下,可以诱导更多的神经干细胞向胆碱能神经元分化.

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