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The differentiation of neural stem cells after tyrosine hydroxylase gene transferring
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF NEUROSURGERY
- Issue:
- 5
- DOI:
- 10.3760/j.issn:1001-2346.2002.05.006
- Key Word:
- 神经干细胞;分化;酪氨酸羟化酶;基因转染;碱性成纤维细胞生长因子
Abstract： 目的探讨神经干细胞(NSCs)转染酪氨酸羟化酶(TH)基因后的分化.方法从胚胎16天Wistar大鼠室管膜前下周围区分离、增殖、鉴定NSCs;将TH基因和缺陷性逆转录病毒载体N2A构建成真核表达质粒,以电穿孔将其转入PA317包装细胞内;收集PA317包装产生的逆转录病毒颗粒,感染体外培养的NSCs,经G418筛选,获得成功转染TH基因的NSCs克隆;分别以0.4ng/ml bFGF和5%胎牛血清诱导转染及未转染TH基因的NSCs分化,比较TH基因转染及不同诱导方式对NSCs分化的影响,同时在分化细胞内检测TH的表达.结果以0.4ng/ml bFGF诱导可使95%以上的NSCs分化为神经元,而5%FBS诱导则大多分化为神经胶质细胞,无论是否转染TH基因,神经元及神经胶质细胞的分化比例不发生改变;TH基因转染后能在神经干细胞的子代细胞内高效、稳定表达.结论 TH基因转染不影响NSCs的分化潜能,TH能在神经干细胞的子代细胞中有效表达;0.4ng/ml bFGF诱导可以促使NSCs分化为神经元.
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