Dental pulp stem cells stimulate neuronal differentiation of PC12 cells

Dental pulp stem cells (DPSCs) secrete neurotrophic factors which may play an important therapeutic role in neural development, maintenance and repair. To test this hypothesis, DPSCs-conditioned medium (DPSCs-CM) was collected from 72 hours serum-free DPSCs cultures. The impact of DPSCs-derived on PC12 survival, growth, migration differentiation investigated. were treated with nerve growth factor (NGF), DPSCs-CM or co-cultured using Transwell inserts for 8 days. number surviving neurite outgrowths the length neurites measured by image analysis. Immunocytochemical staining used to evaluate expression neuronal markers NeuN, microtubule associated protein 2 (MAP-2) cytoskeletal marker βIII-tubulin. Gene levels axonal growth-associated 43 synaptic Synapsin-I, MAP-2 βIII-tubulin analysed quantitative polymerase chain reaction (qRT-PCR). (NGF, brain-derived [BDNF], neurotrophin-3, glial cell-derived [GDNF]) specific ELISAs. Specific neutralizing antibodies against detected study their exact survival outgrowth extension. significantly promoted cell induced confirmed immunostaining. Furthermore, more effective stimulating than live DPSCs/PC12 co-cultures over time studied. morphology similar NGF positive controls; however, stimulation higher what seen NGF-treated markedly reduced addition anti-GDNF, whilst attenuated anti-NGF, anti-GDNF anti-BDNF antibodies. These findings demonstrated that able promote differentiation. NGF, BDNF GDNF involved stimulatory action outgrowth, whereas also had a significant promoting survival. be harnessed as cell-free therapy peripheral All experiments conducted dead animals not sacrificed purpose study. methods carried out accordance Birmingham University guidelines regulations ethical approval is needed.