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目的 :观察成鼠神经干细胞移植入切割海马伞侧海马和正常侧海马后的存活和迁移情况。方法 :取成鼠前脑室下带 (SVZ)组织制成单细胞悬液 ,接种于含 EGF和 b FGF的 DMEM/ F12 (1∶ 1)无血清培养基中 ,待神经球形成后 ,用有限稀释法进行单细胞克隆。将单克隆细胞球进行消化、分离 ,加入 Brd U标记并扩增成大量来源于同一细胞的亚细胞系神经干细胞克隆球。取 SD大鼠 ,切割其右侧海马伞 ,术后 14天 ,将标记有 Brd U的神经干细胞植入两侧海马齿状回中。移植术后 1月 ,取脑冰冻切片 ,作 Nissl染色和 Brd U免疫荧光检测。结果 :Nissl染色片两侧海马齿状回中有许多大小不等的深染细胞沿颗粒下层排列。两侧移植区中的 Brd U免疫荧光标记细胞均从移植区沿海马颗粒下层迁移 ,形成一明显的迁移带。切割海马伞侧海马内的 Brd U免疫荧光亮点密度明显大于正常侧海马内的免疫荧光亮点密度。免疫荧光片 Nissl复染发现齿状回内的深染细胞与 Brd U免疫荧光亮点相对应 ,在切割海马伞侧海马齿状回内有大胞体的神经元样细胞 ,而正常侧较少见。结论 :移植至海马齿状回中的神经干细胞能存活 ,并可沿正常海马齿状回中固有的神经干细胞迁移路径即颗粒下层迁移。切割海马伞侧海马内存活和迁移的神经干细胞明显多于正常侧海马内的神经
OBJECTIVE: To observe the survival and migration of adult rat neural stem cells after transplanted into hippocampus and normal hippocampus of hippocampus. Methods: SVZ tissue was made into single cell suspension and inoculated into DMEM / F12 (1: 1) serum-free medium containing EGF and b FGF. After the formation of neurospheres, Dilution method for single cell cloning. The monoclonal cell spheres were digested, isolated, labeled with BrdU and amplified into a large number of subcellular lines of NSCs derived from the same cell. SD rats were sacrificed and the right hippocampus was cut. After 14 days, the neural stem cells labeled with BrdU were implanted into the dentate gyrus on both sides of the hippocampus. One month after transplantation, brain slices were taken and Nissl staining and BrdU immunofluorescence detection were performed. Results: There were many dark-stained cells in the hippocampal dentate gyrus on both sides of the Nissl stain. BrdU immunofluorescence labeled cells in both grafts migrated from the transplanted subepithelial layer of the hippocampus, forming a clear migration zone. BrdU immunofluorescent bright spot density in the hippocampus of the hippocampus was significantly higher than that in the normal hippocampus. The immunofluorescent Nissl staining showed that the stained cells in the dentate gyrus corresponded to BrdU immunofluorescence bright spots. In the hippocampal dentate gyrus of the hippocampus, there were neuronal cells in the dentate gyrus, while the normal side was rare. CONCLUSIONS: Neural stem cells transplanted into the dentate gyrus of the hippocampus survive and migrate along the migration pathways underlying the neural stem cells, ie, the subgranular, inherent in the hippocampal dentate gyrus. The neural stem cells that survive and migrate in the hippocampus of the hippocampus were significantly more than those in the normal hippocampus