Lin S, Wang Y, Zhang C, Xu J
Department of Orthopaedics, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
J Int Med Res. 2012;40(4):1449-58. doi: 10.1177/147323001204000423.
To investigate the effects of the neurotrophin-3 (NTF3) gene on the survival and differentiation of neural stem cells (NSCs) in vitro and in vivo.
The NTF3 gene was isolated from rats, amplified by polymerase chain reaction (PCR) and subcloned into the lentiviral vector pWPXL-MOD to construct a lentiviral expression vector pWPXL-MOD-NTF3. Reverse transcription-PCR and Western blotting were used to analyse NTF3 mRNA and protein levels, respectively. Adult rats with sectioned tibial nerves received implants of NSCs transfected with either pWPXL-MOD-NTF3 (n=30) or an empty expression vector (n=30). In vitro and in vivo cell differentiation and survival were determined by fluorescence immunohistochemistry.
Expression of NTF3 significantly increased the differentiation of NSCs into cholinergic neurons both in vitro and in vivo. NTF3-expressing NSCs implanted into the tibial nerve also survived longer than cells without NTF3 gene modification.
The NTF3 gene promoted differentiation of NSCs into cholinergic neurons and enhanced neuronal cell survival. These findings may have clinical implications for cell transplantation therapy in patients with nerve injury.
研究神经营养因子3（NTF3）基因在体外和体内对神经干细胞（NSCs）存活及分化的影响。
从大鼠中分离出NTF3基因，通过聚合酶链反应（PCR）进行扩增，并亚克隆至慢病毒载体pWPXL-MOD中，构建慢病毒表达载体pWPXL-MOD-NTF3。分别采用逆转录PCR和蛋白质印迹法分析NTF3的mRNA和蛋白质水平。成年大鼠胫神经切断后，植入用pWPXL-MOD-NTF3转染的神经干细胞（n = 30）或空表达载体（n = 30）。通过荧光免疫组织化学法测定体外和体内细胞的分化及存活情况。
NTF3的表达在体外和体内均显著增加了神经干细胞向胆碱能神经元的分化。植入胫神经的表达NTF3的神经干细胞也比未进行NTF3基因修饰的细胞存活时间更长。
NTF3基因促进神经干细胞向胆碱能神经元分化，并提高神经元细胞的存活率。这些发现可能对神经损伤患者的细胞移植治疗具有临床意义。
