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目的将体外标记的骨髓基质源神经干细胞经单细胞悬液微移植后观察其在兔纹状体的存活、迁移、分化和整合情况,为细胞移植治疗疾病奠定基础。方法分离兔骨髓基质细胞,利用神经干细胞培养基、白血病抑止因子和碱性成纤维母细胞生长因子进行细胞扩增并诱导成骨髓基质源神经干细胞,再经菲立磁和活细胞荧光染料PKH67标记后,采用微移植的方法,通过脑立体定位仪,用微玻璃针将干细胞分别植入兔脑纹状体内。存活1、4、8周后处死动物,组织切片,利用光镜和电镜观察标记细胞在脑内的形态学情况。结果菲立磁标记的兔骨髓基质源神经干细胞经微移植后可在兔脑内纹状体区域存活,移植的干细胞可向周围的脑实质内迁移和整合,迁移细胞沿特定的纹状体结构分布。少量菲立磁标记的干细胞可以分化成神经元。结论骨髓基质源神经干细胞移植后,可在脑实质内存活、迁移、分化和整合,这种细胞可能成为中枢神经系统自体移植的细胞来源。
OBJECTIVE: To observe the survival, migration, differentiation and integration of in vitro labeled bone marrow stromal cells derived from neural stem cells after single-cell suspension micro-transplantation, and lay a foundation for the treatment of diseases by cell transplantation. Methods Rabbit bone marrow stromal cells were isolated and cultured with neural stem cell culture medium, leukemia inhibitory factor and basic fibroblast growth factor to induce bone marrow stromal cells derived from neural stem cells and labeled with phenanthrene and fluorescent dye PKH67 After using micro-transplantation method, through the brain stereotaxic instrument, with micro-glass needle will be stem cells were implanted in the rabbit brain striatum. After 1, 4 and 8 weeks of survival, the animals were sacrificed and the sections were stained. The morphology of labeled cells in the brain was observed by light and electron microscopy. Results Feridex-labeled rabbit bone marrow stromal cells derived from neural stem cells survived in the striatum of the rabbit brain after transplantation. Transplanted stem cells migrated and integrated into the surrounding brain parenchyma, migrating cells along specific striatum structures distributed. Few Philippine-labeled stem cells can differentiate into neurons. Conclusion Bone marrow stromal cells derived from neural stem cells can survive, migrate, differentiate and integrate in the parenchyma of the brain. These cells may become the source of cells for autologous transplantation of the central nervous system.