ABSTRACT
BACKGROUND:In the past, the culture and differentiation of bone marrow mesenchymal stem celsin vitrowere mostly reported in the adult or animal rather than in children. OBJECTIVE: To explore the ability of bone marrow mesenchymal stem cels from children differentiating into neural stem cels and nerve cels. METHODS: Bone marrow mesenchymal stem cels from children were isolated and cultured, and passage 12 cels were cultured in the pre-induction medium (DMEM culture medium containing 10% fetal bovine serum and 1 mmol/L β-mercapto ethanol) and induction medium (DMEM containing 2% dimethyl sulfoxide and 150 μmol/L butylated hydroxyanisole). Expression of nestin and β-tublin III was detected using immunocytochemistry method at 30 minutes and 7 days after induction, while RT-PCR was used to detect nestin mRNA expression at 0, 5.5, 6 days after induction. RESULTS AND CONCLUSION: After combined induction, the cels shrank from round shape to tapered, polygonal or oval shape, and cel processes extended gradualy and became filament-like shape. Interconnected cels formed a network at 6 days after combined induction. The expression of nestin antigen was positive at 30 minutes after induction, while the expression of β-tublin was positive at 7 days. RT-PCR findings showed that positive expression of nestin mRNA was detected at 5.5 hours of induction, and then disappeared at 6 days. These findings show that the combined use of dimethyl sulfoxide and butylated hydroxyanisole can induce bone marrow mesenchymal stem cels from children to differentiate into neural stem cels and nerve cels in vitro.