| ObjectiveBone marrow mesenchymal stem cells(BMSCs)are the stem cells which have the patentied to differentrate into many kinds of cells.In the special condition, BMSCs can differentrate to nerve cells and then like neuron cell with induction. Many kinds of reasons send the facial nerve injury is that oral cavity jaw surface system most common disease. Because of nerve fiber not renewability, it is serious influence treatment and patient prognosis and it is becomes difficult problem which the people face.How to tread the disease and how to improve patient prognosis are the the points at issue to the various countries'scholar widely studies.Nowadays, stem cell transplant treatment and the cell substitution treats become the new way nervous system disease. The study goal is to investigate the culture condition of BMSCs and the method of inducing the diferentiation of BMSCs into neurons in vitro and then identify the neurons.Methods1,BMSCs were isolated from healthy SD rats and purified and cultured.2,Cellsurface antigen was determined by flow cytometry, and BMSCs were identifed by immunophenotype.3,BMSCs were induced by adherent culture and diferentiated into nerve cells and then were divided into control group and inductor group. No inductor was seen in the control group. Firstly, epidermal growth factor and basic fibroblastgrowth factor were added in the inductor group for 6 days, and then brain derived neurotrophic factor and retinoic acid for 6 days. The induced neurons were identified by immunofluorescence staining at days3,6and 9.Results1,The third generation BMSCs appeared fibroblast-like and were positive for the markers CD29,CD44 and CD90 with the positive rates of 98.76%,98.41%,96.58% and negative for typical hematopoietic markers CD34,CD54.2,The BMSCs highly expressed Nestin 83% after 6 day induction in the inductor group,while the control group was negative for this marker.3,The BMSCs expressed microtubule associated protein MAP,NF,NSE after 6 day induction in the inductor group,and most BMSCs diferentiated into neuron-like cells,while compares with the control group has the difference for this marker and cells appeared fibroblast like(P<0.05).Conclusions1,This study utilizes and modifies totaL bone marrow adherence, which is simple and rapid, with good cytoactive,and it is an ideal method to isolate and purify BMSCs.2,The better eftect is acquired with the combination of bFGF and the EGF and better than separated.3,Two kind of prederivative bFGF and EGF density 100ug/1 density preinduction effect for best,can promote BMSCs to the nerve stem cell differentiation.4,After ATRA and the BDNF induction differentiation cell, can the majority of nerve stem cell induction for the neuron type cell.5,The appraisal cell method are plentiful including flow cytometry analysis appraisal BMSCs, Immunofluorescence and Immunohistochemcical appraisal for induction the neuron type cell. 6,The appraisal cell reagent type are plentiful including CD29,CD90,CD45,CD44,CD34,CD90 (appraisal BMSCs),NSE, NF. MAP-2, after NESTIN (appraisal induction neuron type cell).
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