Objective: To explore the effect of Fasudil on the proliferation and survival of bone marrow-derived neural stem cells (BMNSCs) cultured in vitro.
Methods: BMNSCs were derived from C57BL/6 mice aged 3-4 weeks and induced to produce cell spheres in serum-free medium. The cells were detected by immunofluorescence staining with nestin antibody. The effect of Fasudil on the proliferation of BMNSCs was detected by 5, 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) labeling. BMNSCs were cultured under the inflammatory condition created by BV-2 microglia treated with lipopolysaccharide (LPS) and IFN-γ. The mitochondrial membrane potential (MMP) and propidium iodide (PI) fluorescence value of BMNSCs were determined by flow cytometry to explore the protective effect of Fasudil on BMNSCs.
Results: Immunofluorescence staining verified that bone marrow-derived cell spheres were nestin-positive neural stem cells. The treatment of Fasudil significantly decreased the average fluorescence value of CFSE. BV-2 microglias treated with LPS and IFN-γ released more inflammatory factors such as IL-6, TNF-α than PBS treated group. The inflammatory conditioned medium significantly decreased the MMP of BMNSCs and induced more cell death, while the treatment of Fasudil significantly restored the MMP of BMNSCs close to the normal value and reduced cell death in an obvious dose-dependent manner.
Conclusion: Fasudil has significantly promoting effect on the proliferation and survival of BMNSCs in vitro.