[Differentiation of embryonic stem cells into neuronal cells in vitro]

Abstract

Objective: To investigate the probabilities of embryonic stem (ES) cells differentiating into neuronal cells in vitro.

Methods: Mouse ES-D3 cells were cultured on the ES conditioned media without mLIF (mouse leukemic inhibitory factor, mLIF) for 4 days until ES cell aggregations was formed (embryonic bodies, EBs). The EBs were dissociated with trypsin and exposed to retinoic acid (RA) for another 4 days, and then replated onto adhesive substrate. Immunohistochemistry and RT-PCR assay were performed to identify the properties of the differentiated cells.

Results: The treated cells expressed neuronal proteins such as MAP-2 and GFAP. RT-PCR assay further identified that the induced cells expressed transcripts for neural associated genes including GABA-gamma subunit receptor, brain factor-1 and tyrosine hydroxylase (TH).

Conclusion: ES cells can be successfully induced into neuronal cells expressing multiple properties of neurons in vitro, which could provide resourceful materials for neural transplantation.