Objective: To study the induction of Xiangdan Injection (XDI) on the committed differentiation into neuron from rMSCs.
Methods: The 5th passage of rMSCs were pretreated with alpha-MEM and 10 microg/L basic fibroblast growth factor (bFGF) for 24h, and treated with the serum-free induction media (D/F12 + N2 + bFGF (10 microg/L) and 1% - 5% XDI). The differentiated cells were observed with phase-contrast microscopy and detected the expression of several specific proteins, such as neurofilament-200 (NF-200), neuron-specific enolase (NSE), microtubule-associated protein-2 (MAP-2) and glial fibrillary acidic protein (GFAP) with immuno-cytochemistry and semi-quantity reverse transcriptase-polymerase chain reaction (RT-PCR).
Results: At 3h of induction, rMSCs gradually turned into the neuron-like cells in the induction groups. The induction rate can reach 83.5%+/-3. 8% at 6 - 8 h of induction. The percentages of responsive cells positive for NSE,NF-200 and GFAP were 82.9%+/-2.98%, 84.1%+/-4.45% and 3.49%+/-1.67% respectively. RT-PCR showed that the relative level of expression for NSE and MAP-2 was much higher than that of GFAP (over 33 fold) (P <0. 01). The unresponsive cells in the induction group and those cultured cells in the control were shown negative for all of the for- mer detected markers.
Conclusions: The rMSCs could be differentiated into neuron-like cells in vitro with the induction of XDI in the free-serum media.