Directed differentiation of mouse embryonic stem cells into neuronal cells induced by icaritin in vitro

Abstract

To evaluate the inductive effects of icaritin (ICT) on the directed differentiation of mouse embryonic stem (ES) cells into neuronal cells in vitro. ES cells were cultured with embryoid body (EB) formation cultures, ICT in different concentrations was added in the cultural media and the cells were harvested in several differentiation phases. The expression spectrums of neuronal cell-specific genes and proteins were verified by semi-quantitative RT-PCR and immunocytochemistry analysis, respectively. Differentiation of neurocyte phenotype from ES cells was promoted by ICT in a concentration-and time-dependent manner. ICT at 10(-7)mol/L significantly enhanced the differentiation toward neuronal cells, and up to 80 % of EBs outgrowth in d 8+8 incubation. The gene expressions of beta-tubulin III in neuron and GFAP in glial cells were detected in neuronal cell phenotype derived from EBs. Furthermore, nestin was detected in precursor cells, beta-tubulin III and GFAP were detected in the generated precursor neurocytes immunocytochemically. Directed differentiation of neurons is facilitated by ICT in EB formation culture, which is associated with the expression of developmental-dependent gene and protein.