胞嘧啶脱氨酶基因修饰神经干细胞及其表达的离体实验研究

目的 探讨胞嘧啶脱氨酶(cytimidine deaminase，CD)基因修饰神经干细胞及其基因表达。方法 通过构建真核表达质粒pCMVCD，限制性内切酶消化鉴定后，采用Lipofectamine 2000脂质体介导法转染新生大鼠室管膜下区神经干细胞(Neural stem cells，NSCs)，G418筛选阳性克隆，加入不同浓度的5-氟胞嘧啶(5-Flourocytosine，5-FC)，MTT比色法测定NSCs的生存率。结果 本实验成功地培养并鉴定了神经干细胞，并将CD基因成功地转染了神经干细胞，G418阳性NSCs对低浓度5-FC高度敏感。结论 CD基因修饰神经干细胞的离体实验研究为干细胞治疗研究提供依据。

Study of neural stem cells modified by cytosine deaminase gene and its expression in vitro

Objective To investigate neural stem cells (NSCs) modified by cytosine deaminase (CD) gene and its expression in vitro . Methods Eukaryotic expression plasmid pCMVCD was constructed, and identified by restriction endoenzyme digestion. CD gene was transfected into NSCs from new born Wistar rats using Lipofectamine2000. Positive clones (named NSCs/CD cells) were screened by G418 presence. 5 Fluorocytosine (5 FC) administration of different concentrations were incubated with NSCs/CD cells. NSCs/CD cells viability ratios were measured by MTT assay. Results NSCs cell culture and identification were carried out, transduction of CD gene was also successfully proceeded. Gene transfer made G418 positive NSCs (NSCs/CD cells) highly sensitive to 5 FC. Conclusion Study of NSCs modified by cytosine CD gene and its expression in vitro plays an important role in the studies of stem cell therapy in vivo .