大鼠羊膜上皮细胞体外向类神经干细胞诱导分化及其移植后对视神经再生的作用

发布时间：2018-01-18 16:06

本文关键词：大鼠羊膜上皮细胞体外向类神经干细胞诱导分化及其移植后对视神经再生的作用　出处：《第二军医大学》2010年硕士论文　论文类型：学位论文

【摘要】： 视觉系统一些疾病会造成视神经损伤而引起患者失明,目前临床上尚无良好的治疗方法,而视神经再生的研究将为治疗这些疾病提供实验依据。本研究通过体外培养妊娠晚期大鼠羊膜上皮细胞,予以免疫细胞化学、RT-PCR等方法鉴定后,应用不含血清的神经干细胞培养基诱导大鼠羊膜上皮细胞向类神经干细胞方向分化,并予以免疫细胞化学、RT-PCR等方法鉴定。然后,将Hoechst33342荧光标记的正常羊膜上皮细胞与诱导后的羊膜上皮细胞分别移植入大鼠受损视神经中,应用Hoechst33342荧光追踪移植细胞、视网膜HE染色和尼氏染色计数视网膜节细胞数量、视神经HE染色、免疫组织化学染色等方法初步探讨细胞移植后对视网膜节细胞的存活和视神经轴突再生所产生的作用。结果表明,应用不含血清的神经干细胞培养基可诱导大鼠羊膜上皮细胞向类神经干细胞方向分化；诱导前后的羊膜上皮细胞移植入受损视神经后,能在损伤部位大量存活并向损伤区两端迁移,能提高视网膜节细胞存活率,增加远侧段视神经的细胞数量,促进视神经轴突再生。相对来说,诱导后的羊膜上皮细胞较诱导前的细胞移植对视神经修复作用更强些。从而提示我们诱导后的羊膜上皮细胞经移植后能促进视神经损伤的修复,为临床修复视神经提供新思路。
[Abstract]:Some diseases of the visual system can cause blindness in patients with optic nerve injury, and there is no good treatment in clinic. The study of optic nerve regeneration will provide experimental basis for the treatment of these diseases. In this study, amniotic epithelial cells were cultured in vitro and identified by immunocytochemistry and RT-PCR. Rat amniotic epithelial cells were induced to differentiate into neural stem cells (NSCs) by using serum-free neural stem cells (NSCs) medium, and were identified by immunocytochemistry and RT-PCR. Hoechst33342 fluorescent labeled normal amniotic epithelial cells and induced amniotic epithelial cells were transplanted into the injured optic nerve of rats. The number of retinal ganglion cells and optic nerve HE staining were counted by Hoechst33342 fluorescence tracing, HE staining and Nissl staining of retina. The effects of cell transplantation on the survival of retinal ganglion cells and the regeneration of optic nerve axons were preliminarily investigated by immunohistochemical staining. The differentiation of rat amniotic epithelial cells into neural stem cells was induced by using neural stem cell culture medium without serum. After transplantation of amniotic epithelial cells into the injured optic nerve before and after induction, they can survive and migrate to both ends of the injured area, which can improve the survival rate of retinal ganglion cells and increase the number of distal optic nerve cells. Promote axonal regeneration of the optic nerve. Relatively speaking. The effect of induced amniotic epithelial cells on optic nerve repair is stronger than that of pre-induced amniotic epithelial cells, which suggests that the induced amniotic epithelial cells can promote the repair of optic nerve injury after transplantation. To provide a new idea for clinical repair of optic nerve.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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