Experiment Study On Brain-derived Neurotrophic Factor To Promote The Differentiation Of Muscle-derived Stem Cells Into Neurons-like Cell In Vitro

Objective:1. To investigate the methods about the segregation, purification, cultivation and identification of Muscle-derived Stem Cells (MDSCs) in vitro, especially, to study the cells' biological characters. These studies are preparations for the succeeded experiments.2. To assess the different dosage of BDNF to induce Muscle-derived Stem Cells(MDSCs) into neurospher-like cells, and to examine the expression of neurospher-like cells after being transfected. According to those, we can provide experimental bases for the repair of bone defect by utilizing MDSCs.Method:1. The rats' MDSCs were acquired by enzyme digestion. By two-step method, the rat muscle was digested with type XI collagen and trypsin, and the isolated cell suspension was purified the millipore nylon net filter. The proliferation ability of MDSCs was analyzed through studying growth curve, and the obtained cells were identified with cell immunocytochemical technique.2. Nestin molecule of these cells was inspected after they were cultured in non-adherent condition with bFGF and EGF. After the different dosage of BDNF to induce MDSCs, the expression of NSE and GFAP was detected in these cells after they were induced in adherent cultural condition with different dosage of BDNF. Result:1. The isolated MDSCs were small-cycloid or short-fusiform-shaped. And the isolated cell suspension was purified the millipore nylon net filte (11um, 20um, 30um, 41um) under the growth medium, MDSCs trend to proliferation, however, the cells trend to differentiate. They showed desmin (+) and CD₃₄ (+) by cell immunocytochemical technique.2. After being passaged, most of cells adherenced grow, a few of cells suspension lumping. Suspension cells grew quickly, neurosphere-like cell agglomerates were gained during 7-15d. The different dosage of BDNF to induce the neurosphere-like cell agglomerates, the cells' morphous was changed. They showed GFAP (+) by cell immunocytochemical technique.Conclusion:1. MDSCs in vitro showed strange proliferative and differentiated ability, and the isolated cell suspension was purified the millipore nylon net filter. We could modulate MDSCs' proliferation and differentiation by regulation the serumal concentration. As the target cells, it was a good choice for tissue engineering and gene therapy.2. The different dosage of BDNF to induce Muscle-derived Stem Cells (MDSCs) into neurospher-like cells. MDSCs could applicate to repair born coloboma, muscle atrophy, myocardial infarction and stress incontinence, moreover, it could treat the disease of nervous system.