胚胎干细胞源性视网膜前体细胞在rd鼠视网膜下腔移植后的分化  

Differentiation of embryonic stem cell derived retinal precursor cell after transplanted into subretinal space in rd mouse

作　　者：陶靖[1] 葛坚[2] 

机构地区：[1]首都医科大学附属北京同仁医院,北京同仁眼科中心,北京市眼科学与视觉科学重点实验室,北京市100730 [2]中山大学中山眼科中心,广东省广州市510060

出　　处：《眼科新进展》2009年第7期485-487,491,共4页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30171001;30672275);国家重点基础研究发展计划(973计划)项目基金资助(编号:2007CB512200)~~

摘　　要：目的研究绿色荧光蛋白标记的胚胎干细胞(green fluorescent protein-embryonic stem cells,GFP-ESC)源性视网膜前体细胞在rd小鼠视网膜下腔移植后的存活分化情况。方法将GFP-ESC源性视网膜前体细胞移植到伴有视网膜感光细胞变性和继发性视网膜神经节细胞变性的rd小鼠视网膜下腔,移植后1周、4周、8周取材行免疫荧光细胞化学检测,观察移植细胞存活、分化情况。对移植细胞进行核型分析,并接种裸鼠眼内观察是否成瘤,评价其安全性。结果GFP-ESC源性视网膜前体细胞移植眼内可存活、整合到宿主变性视网膜层间。1周时GFP阳性细胞移行于外层视网膜并呈视杆细胞标志抗原rhodop-sin阳性;4周时整合于内层视网膜,共表达成熟神经元标志抗原神经丝蛋白-200、微管相关蛋白-2,突触标志抗原synaptophysin检测阳性;8周时移至邻近玻璃体腔形成节细胞样结构,表达节细胞标志抗原Thy1.1;移植细胞维持正常二倍体核型,术后8周未见排斥反应及瘤性增殖。结论GFP-ESC源性视网膜前体细胞在变性视网膜中可存活、整合,并有优先向变性的感光细胞及神经节细胞类型分化的倾向,有望为青光眼等视神经视网膜病变的细胞移植治疗提供安全有效的种子细胞。Objective To investigate the survival and differentiation of green fluorescent protein-embryonic stem cell （GFP-ESC） derived retinal precursor cells after transplanted into subretinal space in rd mouse. Methods GFP-ESC derived retinal precursor cells were transplanted into subretinal space in rd mouse, which was with primary rob cell degeneration and secondary ganglion cell degeneration. The survival and differentiation of transplanted cells were detected with immunofluorescent cyto- chemical method at 1 week, 4 weeks and 8 weeks after transplantation, respectively. Transplanted cells were taken with karyotype analysis, and inoculated nude mouse was intraocular observed to have tumor or not. The safety was evaluated. Results GFP- ESC derived retinal precursor cells could survive and integrate into the degenerated layer of host retina. At 1 week after transplantation, GFP positive cells migrated into outer layer of retina and expressed positive rob cell-antigen rhodopsin. At 4 weeks after transplantation, GFP positive cells integrated into inner layer of retina and expressed mature neurons-antigen neurofilament protein-200 and microtubule associated protein-2 as well as positive synapse-antigen Synaptophysin. At 8 weeks after transplantation, GFP positive cells moved to adjacent vitreous space, formed nodal cell liked structure, and expressed nodal cell-antigen Thyl. 1. The transplanted cells maintained diploid kar- yotype, and there was no rejection or tumor proliferation at 8 weeks after transplantation. Conclusions GFP-ESC derived retinal precursor cells can survive and integrate in degenerated retina, and have the tendency to preferentially differentiate into degenerated photoreceptor cells and ganglion cells. They have the potential to provide safe and effective seed cells in cell transplanted treatment of neural retinopathy such as glaucoma

关 键 词：胚胎干细胞 视网膜前体细胞 定向分化 神经再生 

分 类 号：R777.2[医药卫生—眼科]