Abstract
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.