【作者】 尹成玉；

【导师】 王斌；

【作者基本信息】 青岛大学 ， 病原生物学， 2011， 硕士

【摘要】 目的观察间充质干细胞治疗视神经损伤的安全性和疗效。方法以MEM/12培养基,采用淋巴细胞分离液间接分离法,分离间充质干细胞(MSCs),以巾壁培养法进行原代培养和纯化。倒置显微镜下观察生长状态和形态学变化,共聚焦显微镜下观察细胞表面抗原CD34,CD44、CD29免疫组化染色的鉴定,分离获得间充质干细胞；搜集2010年12月至2011年9月来我院住院的视神经损伤患者23人,其中男15人,女8人,年龄在5-77岁之间,平均年龄30.1岁,其中原发性视神经萎缩12人,遗传性Lebers病5人,视神经发育不良者1人,视隔发育不良者2人,视网膜色素变性者3人。治疗前后进行各项指标检测,包括谷丙转氨酶、谷草转氨酶、尿素氮、肌酐、血糖、甘油三酯、胆固醇、乙肝表面抗原、凝血酶原时间、部分凝血酶原时间、甲胎球蛋白、癌胚抗原检查,眼科检查视力、眼压、测杯盘比等,采用经分离获得的干细胞对收集的23位视神经损伤患者分四个阶段进行治疗,每次给药间隔3-5天,每次给予2-4*107个干细胞。对视神经损伤患者治疗前后检测闪光型视觉诱发电位的进行对比,分析间充质干细胞对视神经损伤的治疗效果。结果荧光显微镜下可见均强烈表达CD44,而CD34无表达。表明MSCs表达间充质细胞表面抗原而不表达造血细胞、祖细胞及内皮细胞表面抗原,这与骨髓来源的MSCs免疫组化结果一致。干细胞治疗后后实验室指标、眼压、测杯盘比无显著性差异,视力、闪光型视觉诱发电位的潜时、波幅变化明显。结论成功获取间充质干细胞；间充质干细胞移植治疗视神经损伤患者的过程安全可靠；间充质干细胞移植治疗视神经损伤患者视力、闪光型视觉诱发电位均有改善。更多还原

【Abstract】 Purpose Observe the safety and curative effect of mesenchymal stem cell therapy for optic nerve injury (ONI).Methods Use MEM as culture medium, by the lymphocyte separation liquid indirect separation, separate mesenchymal stem cells (MSCs) by adherent culture for primary culture and purification. Observe cell growth state and morphological changes under inverted microscope, and observe immunohistochemical stains appraisal of cell surface antigen CD34, CD44, CD29, isolate mesenchymal stem cells by focusing microscope. Collected in December 2010 to September 2011 ONI to our hospital 23 patients in our hospital between Dec 2010 and Sep 2011,15 male and 8 female, between 5 and 77 years, and average age was 30.1 years old, with 12 primary optic nerve atrophy (ON A) patients,5 Leber Hereditary Optic Neuropathy (LHON) patients,1 Optic Nerve Hypoplasia (ONH) patient,2 Septo Optic Dysplasia (SOD) patients, and 3 Retinitis Pigmentosa (RP) patients. We detected every indicator before and after treatments, included ALT, AST, BUN, creatinine, blood sugar, TG, cholesterol, HBsAg, PT, partial thromboplastin time (APIT), AFP, CEA and ophthalmologic examination:visual acuity(VA), intraocular pressure(IOP), cup to disk ratio. Using isolated stem cells treated for collected 23 optic nerve injury patients in 4 treatments, each treatment gave 2-4×10’stem-cells apart 3 to 5 days. By comparing the Flash visual evoked potential(F-VEP)results before and after treatments for ONI patients, analyse the treatment effect of MSCs.Result Strong expression of CD44, and no expression CD34 under fluorescent microscope are. Show that MSCs express mesenchymal cells surface antigen and do not express hematopoietic cells, progenitor cells or endothelial cell surface antigen. It was accordant with bone marrow-derived MSCs immunohistochemical results. There was no significant difference in laboratory indicators, IOP and cup to disk ratio, however, there were marked difference in VA, latency and amplitude of F-VEP before and after treatments. Conclusion Succeed in obtaining mesenchymal stem cells. The procedure of mesenchymal stem-cells treatment for ONI patients is safe and reliable. Mesenchymal stem-cells transplantation is able to improve ONI patients’VA, F-VEP.更多还原