Observation Of SD Rat Spiral Ganglion Cells And Bone Marrow Mesenchymal Stem Cells Co-culture

Objective taking SD rat the original spiral of ganglion cells and bone marrow mesenchymal stem cells indirect trained, Observation of bone marrow stromal whether induced into nerve cells,for treating neurological deafness seek new treatments.Method First, the original training of bone marrow mesenchymal stem cells:the bone marrow direct stick wall of SD rats BMSCs training,taking the third generation cells for flow cytometric detection and immune cells chemical dyeing. Second, the original training of spiral ganglion cells:taking1-3days of SD rats training SGCs in vitro,taking the third day cells in the immune cells chemical dyeing to identify with neuron-specific enolase (NSE).Third, adopting the indirect method of cultivation,taking spiral of ganglion cells with the third generation bone marrow mesenchymal stem cells to cocultivate in an approximately1:1ratio,observating cells everyday.Taking5-7days of bone mesenchymal stem cells in immune cells chemical dyeing with NSE,observing whether there are neurons-like cell with dyeing positive.Result First, the method of enzyme digest after training spiral of ganglion cells24hours,Most of cells stick to wall, and send forth short axon,the axon gradually become long after48-72hours, cells is oval or round, axon is slender, and often interweaves mesh,after training six to fourteen days, the cells gradually reduce obviously, some cells close to apoptosis.while the method of tissue mass, after inoculating tweenty-four hours, their shap are similar to the cell with enzyme digest, but they survival longer. Second,The original of the training of the generation of bone marrow stromal stem cells stick wall after24h to48h, they growth colony-liking, the cells are short spindle type, many Angle type, etc. after training6days, the cell proliferate quickly, after cultivating11days, cell growth of the bottle80-90%, it is time to generate the cell, the second generation of cell of proliferation speed are faster compared to the first generate, training to the third generation, the cells shape appear "spindle",and show clear vortex sample of growth. Flow cytometric detection indicate that the surface antigen CD34of hematopoietic stem cells and endothelial cells express appare negative,and the surface antigen CD44of mesenchymal stem cell surface appare positive, immunohistochemical is positive, so combining cells form analysis,these cells according with the characteristics BMSCs rats. Third. in co-culture group, bone marrow stem cells began to contractive decrescent, elevated,the tuber increase, some cells are appear star, compared to the control group spiral ganglion cells in co-cuturer group are more longer, the number of the cells decrease after six days later, and the control group, the spiral of ganglion cells day began to appear to reduce after four days later, after continue to cultivate the cells,we found that the speed of the number and the shorter of axons of spiral ganglion cells in co-culture group are slower than the contron group. after training15days, the number of spiral ganglion cells in co-culture group reduce obviously, in the control group, the spiral of ganglion cells close to apoptosis. taking the7day of bone marrow stromal stem cells of co-culture group for immune cells chemical dyeing, a part of cells colour, dyeing time is about5minutes, the positive rate is about15%, in the first control group,bone marrow, mesenchymal stem cells almost negative dyeing。Conclusion first,it is Successful to culture the spiral of ganglion cells in vitro, the spiral of ganglion cells of tissue mass method survival longer,compare to the method of enzyme digestion. Second.the method to get SD rats bone marrow mesenchymal stem cells with attachment culture purely is simple and feasible.third.the environment provided by spiral of ganglion cells in the process of growing have stimulative induced effect to bone marrow stromal stem cells.