[Study on inducing differentiation of mouse embryonic stem cells into neuronal in vitro].

OBJECTIVE: To investigate the probabilities and properties of embryonic stem (ES) cells differentiating into neuronal cells in vitro by retinoic acid. METHODS: Mouse ES D3 cells were cultured by means of handing drop and suspending on the ES conditioned media for 3 days and 1 day until ES cell aggregations was formed (embryonic bodies, EBs). The EBs were exposed to retinoic acid (RA) for another 4 days. Immunohistochemistry, RT-PCR assay and the fluorescence intensity changes of calcium after Glu stimulation were performed to identify the characteristics and differentiation efficiency of the differentiated cells. RESULTS: Expression of nestin protein was detected by immunohistochemistry after 96 h induction and the nestin positive cells were (53.49 +/- 6.02) %. RT-PCR assay further identified that the induced cells expressed transcripts for neural associated genes including nestin, glutaminase and Brn-3. And the induced cells have the same character with neuronals after Glu stimulation. CONCLUSION: ES cells can be successfully induced by retinoic acid into neuronal cells expressing multiple properties of neurons in vitro, and these cells have the similar characters with the neuronal derived in vivo.