Pap Promote Bone Marrow Mesenchymal Stem Cells Into Neuron-like Cells Into Experimental Studies,

Objective: In this paper, biological experimental techniques, research polypeptide (Pilose antler polypeptides, PAP) to promote inducer inducing bone marrow mesenchymal stem cells (bone mesenchymal stem cells, BMSCs) into neuron-like cell differentiation. Through the effects of different concentrations of PAP in vitro induced BMSCs promote neuron-like cells to different periods, so as to identify the optimal induction program. Clinical application for the PAP to provide more and better experimental basis for spinal cord injury to provide a new therapeutic approach.Methods: In aseptic conditions, taking healthy bone marrow of male SD rats by density gradient centrifugation to obtain nuclear cell layer, was inoculated in cell culture flask, placed in the cell incubator. The medium was changed to go through unless passage BMSCs, are purified BMSCs. Determined by the cell cycle and identification of whether the method has identified a number of differentiation, the cells are indeed BMSCs. For bFGF + EGF-based with different doses of PAP of the culture medium for the experimental group, and bFGF + EGF group based medium and standard culture medium as a control study group. Used to observe the 7th day showed nerve cell morphology, the number of neuron-like cells further increased, and the number of cells connected into a network between processes. 1,5,7 d after induction, respectively, under inverted microscope to examine cell morphology, and in the first 7 days was used to observe cell morphology, then immunostaining in the identification of neuron-specific enolase (NSE), neuron specific markers neurofilament (NF), glial fibrillary acidic protein (GFAP) expression.Results: Before induction, rat BMSCs to the main spindle, and some were round, polygonal, a small number of short processes. (L) in the induction of 1d, the inverted microscope visible part of the cell body contraction, refraction, extended processes, some contraction of cell body, forming a triangle or a simple bipolar or multipolar cells. 5d longer processes after the formation of growth cone-like end of the enlargement, showing neuron-like morphology. 7d after the increased number of neuron-like cells, cell processes extend. 7d induced expression after NF, NSE, but cells did not express GFAP. (2) to bFGF + EGF + PAP for the induction agent BMSCs gradually after the induction of neuron-like cells, and showed neuron-like cell morphology, the induction of immune staining after 7d identified NSE, NF is positive, GFAP negative expression.Conclusion:1. PAP can promote the induction of BMSCs into neuron-like cells, differentiation is: BMSCs-neural stem-like cells - neuron-like cells.2. BFGF + EGF + PAP supernatant can induce BMSCs to differentiate into neuron-like cells, which the dosage of 40ug PAP induced the best, better than only using the inductor group, and to some extent, induction dose by PAP effects.