Modification of the Neurotrophin-3 Gene Promotes Cholinergic Neuronal Differentiation and Survival of Neural Stem Cells Derived from Rat Embryonic Spinal Cord In Vitro and In Vivo

Abstract

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.