Fasudil may induce the differentiation of bone marrow mesenchymal stem cells into neuron-like cells via the Wnt/β-catenin pathway

Bone mesenchymal stem cells (MSCs) are an excellent donor graft source due to their potential for self-renewal and multidirectional differentiation. However, it is difficult to obtain high quality MSCs and to induce them to differentiate into neuron-like cells. Fasudil, a Rho kinase inhibitor, exhibits therapeutic potential in spinal cord injuries and stroke. The present study investigated the effect of fasudil on the differentiation of MSCs into neuron-like cells. MSCs were obtained from rat femur marrow, expanded in culture medium, and used at the third passage for subsequent experiments. MSCs were pre-induced with 10 ng/ml basic fibroblast growth factor (bFGF) for 24 h, which was followed by induction with fasudil. A control untreated group and a group treated with fasudil + XAV939, a Wnt/β-catenin pathway inhibitor, were also used in the present study. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR), western blot analysis and immunofluorescence staining were performed in order to detect neuron-specific markers, including neuron-specific enolase (NSE), nestin and neurofilament-M (NF-M). Following induction with fasudil, neuron-like cell morphology was observed. In the fasudil + XAV939 and control groups, no obvious changes in cell shape were observed. The results of RT-qPCR, western blot analysis and immunofluorescence staining indicated that expression of the neuron-specific markers NSE, nestin and NF-M was detected in the fasudil group. The differentiation of MSCs into neuron-like cells induced by fasudil was eliminated when the Wnt/β-catenin pathway was inhibited. The present study demonstrated that fasudil may induce MSCs to differentiate into neuron-like cells, however further studies are required to determine the specific mechanisms involved in the effect of fasudil on the Wnt/β-catenin pathway. In addition, further research is required to examine the functional characteristics of the induced neuron-like cells, in order to establish their suitability for clinical treatments in the future.