Expternal Study Of Adult Rat Bone Marrow Stromal Cells Differentiate Into Nerve Cells

Objective: Bone marrow stromal Cells(BMSCs), as one kindof adult stem cell, are characterized by their abilities of certain heightproliferation, self-renewal and multipotent differentiatiate into various lineagesof tissue cells. The source of BMSCs have easily and conveniently to get,technical conditions are uncomplicated for cultivation in vitro and amplification,besides, BMSCs have the ability of entirely possible to nervous system celldifferentiation by using technologies such as induced culture passage. Thisstudy was base on the establishment of BMSCs isolation, cultivation,purification and amplification system in vitro, devoted on the biologicalcharacteristics of adult rat bone marrow-derived BMSCs and the potential ofBMSCs on differentiating into nerve cells. Methods: In our study, the wholebone marrow adherent cultivation was adopted, based on the characteristics ofadherent BMSCs cells fast; non-adherent cells were removed by replacement ofmedium at the early vaccination, medium was changed repeatedly for purified.After the primary and continuous cell cultivation, biological features andultrastructure were observed under phase contrast microscope, growth curveswere drawn. R-type dyeing were adopted to observe cell morphology while thespecial mark antigen of CD44was detected by immunocytochemistry. On thebase of above, BMSCs were induced to differentiate into neural cells by basicfibroblast growth factor (bFGF) and epidermal growth factor (EGF), the changes of cell morphologic observed, the expression of β-TubulinIII and glialacidic fibrillary protein (GFAP) were observed as neurons specific indicators byimmunocytochemistry. Results:1. Adherent BMSCs were isolated and purifiedbased on the whole bone marrow cultivation with the original generation ofvisible many mixed blood cells growing slowly. Extended14days of70-80%,after three generation cells tended to be consistent. The cell morphology withR-type dyeing and the expression of special mark antigen CD44suggestedpositive detected by immunocytochemistry.2. In this study we used bFGF andEGF to induce BMSCs differentiate into neuron-like cells, the change in formof cells was observed and the expression of special mark antigen β-TubulinIIIand GFAP also suggested positive detected by immunocytochemistry.GFAP-positive cells was significant difference (86±8in one field) from thecontrol group (P <0.05), beta-TubulinIII-positive cells had a significantdifference (78±6in one field) compared to control group (P <0.05), either.Conclusion：1. In the experiment, isolation and purification of whole bonemarrow adherent, marrow stromal cells were easily to be obtained, cells havethe advantages of high survival rate and proliferate stabilize. The originalgeneration showed pleomorphism, conformity in amorphous appeared at thethird generation.2. BMSCs could differentiated into neuron-like cells inducedby bFGF and EGF, observation of differentiation in the cell phases, cytoplasmicand nuclear morphology showed that neural cell-like structure such as cellsgrow synapses could be interconnected into form a neural network, meanwhile, expression of beta TubulinIII and GFAP was obviously clear on the cellmembrane.3. Multiple differentiation potential cells of BMSCs did exist in theadult rat bone marrow, it was easily for amplification in vitro, differentiated intonerve cell under certain conditions, and BMSCs was likely to be a goodpotential cell type in the cell transplantation therapy of neurological diseases.