Objective       To determine the role of β-catenin gene in the differentiation of spinal cord derived neural stem cells.       Methods        After the spinal cord tissue was harvested from fetal rats at embryonic day 14

single cell cloning technique was used to separate and amplify neural stem cells (NSCs)

which were identified with immunofluorescence assay. Then the cells were divided into 3 groups

that is

separately cultured NSCs (blank group)

adenovirus vector AdGFP infected NSCs (GFP group)

and Adβ-catenin adenovirus vector infected NSCs (β-catenin group). Cell infection efficiency was observed by fluorescence microscopy. The mRNA expression of neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) was detected by RT-PCR

and the locations of neurons nucleoprotein (NeuN) and GFAP were observed by immunofluorescence assay. Western blotting was used to detect the protein expression of NeuN and GFAP.       Results       The cells derived from fetal rat spinal cord tissue had the ability of continuous differentiation and cloning

and started to form a small number of cell clusters. The cells were irregular in shape after mechanical separation. The majority of early separated cells were positive to Nestin antigen. The mRNA expression level of NSE was significantly higher in β-catenin group than blank group and GFP group (P0.05)

and that of GFAP was lower than the other 2 groups (P0.05). Immunofluorescence assay showed that the expression of NeuN was significantly higher in β-catenin group than blank group and GFP group (P0.05)

and it was mainly located in the nuclei

but the expression of GFAP was lower (P0.05). Western blot results showed that NeuN was highly expressed in β-catenin group than blank group and GFP group

but GFAP was lowly expressed in β-catenin group (P0.05).       Conclusion       β-catenin promotes the differentiation of spinal cord derived NSCs into neuron.