2005•Chinese Journal of Neurosurgical Disease ResearchRequires access
Biological characteristics of Schwann cells transfected with NT-3 gene
Yao Yuan
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Abstract
Objective To study the biological characteristics of Schwann cells (SCs) transfected with neurotrophin -3 (NT-3) gene in vitro.Methods The NT-3 cDNA was cloned by reverse transcription polymerase chain reaction (RT-PCR) method from human liver tissue, and its eukaryotic expression vector pIRES2-EGFP-NT-3 was constructed and identified by PCR and double-enzyme digestion. The pIRES2-EGFP-NT-3 vector was transfected into SCs by using lipofectamine. The SCs with enhanced green fluorescent protein (EGFP) were observed by fluorescence microscopy. The expression of NT-3 in transfected SCs was determined by immunohistochemical method and grey density measure.Results The eukaryotic expression vector pIRES2- EGFP -NT-3 was successfully constructed. The immunohistochemical staining showed that the NT-3 immunoreactivity was observed both in SCs transfected with pIRES2-EGFP-NT-3 and pIRES2-EGFP, but the latter was weaker.Conclusion The current study provides a new possible way for treatment of the injured facial nerve in the future.