Lhx8促进海马神经干细胞向胆碱能神经元分化

Lhx8 promotes the differentiation of hippocampal neural stem cells into cholinergic neurons in vitro

目的构建大鼠Lhx8全长结构基因真核表达载体。方法 PCR法扩增Lhx8全长结构基因,经EcoRI和BamHI双酶切后连接至真核表达载体pEGFP-C3中,重组质粒pEGFP-C3-Lhx8经测序鉴定后,采用脂质体转染的方法转染至体外培养的海马神经干细胞中。结果测序鉴定表明,重组质粒pEGFP-C3-Lhx8-EGFP构建成功,该重组质粒转入体外培养的神经干细胞后,质粒转染组中ChAT阳性的细胞数较阴性对照组明显增多(P<0.05)。结论重组质粒pEGFP-C3-Lhx8-EGFP构建成功,Lhx8能够促进NSCs向ChAT阳性的细胞分化。

Objective To construct eukaryotic expression vector containing full length cDNA of rat Lhx8 gene. Methods Full length cDNA of rat Lhx8 gene was acquired by PCR method and digested by EcoRI and BamHI. The digested gene fragment was inserted into eukaryotic expression vector pEGFP-C3 and identified by DNA sequencing. The recombinant plasmid was transfected into hippocampal neural stem cells （NSCs） by liposome. Results DNA sequencing demonstrated that the insertion fragment in recombinant plasmid was right, and recombinant plasmid pEGFP-C3-Lhx8 was transfected into NSCs. After 7 days, the number of ChAT positive ceils in plasmid transefection group was significantly higher than the negative control group. Conclusion Eukaryotic expression vector pEGFP-C3- Lbx8 is constructed successfully, and Lhx8 promotes the differentiation of NSCs into ChAT positive cells in vitro.