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1.
Wang W  Ma M  Zhang H  Bai F 《中华眼科杂志》2000,36(5):375-377,I024
目的 以遗传性视细胞变性动物RCS鼠为受体,Wistar鼠为供体,观察纯视锥、杆细胞移植术后RCS鼠视网膜外层超微结构的变化。方法 应用视网膜板层切削技术或准分子激光技术制备供体层状视细胞;视网膜板层外路移植技术,获得视网膜移植动物模型。术后2周及4周时处死动物,眼球常规切片后于光镜及透射电镜下观察受体视网膜。结果 移植的视细胞大多成层排列于受体鼠的视网膜色素上皮层及内颗粒层之间。重新出现的外网状  相似文献   

2.
大鼠骨髓间充质干细胞视网膜下移植的短期观察   总被引:2,自引:0,他引:2  
目的探索骨髓间充质干细胞(MSCs)视网膜移植的可行性。方法采用贴壁筛选法分离、培养SD大鼠骨髓MSCs,将溴脱氧尿嘧啶(BrdU)标记的大鼠MSCs悬液经玻璃体腔注入大鼠视网膜下腔。术后14d处死大鼠,取新鲜眼球,连续冰冻切片,经BrdU单抗、FITC标记二抗和Rhodopsin单抗、Cy3标记二抗免疫荧光双重标记后,荧光显微镜下观察。结果荧光显微镜下可见移植后的MSCs主要分布于视网膜色素上皮层和视锥、视杆细胞层,未表达视网膜光感受器细胞的特异性抗原视紫红质。结论移植14d后MSCs可在视网膜下腔存活,主要分布于视网膜色素上皮层和视锥、视杆细胞层,未分化成视网膜光感受器细胞。  相似文献   

3.
目的 研究小鼠实验性视网膜脱离后光感受器细胞的凋亡情况。 方法 将成年C57Bl/6J小鼠36只分为2组:实验组小鼠18只左眼视网膜下注射1.4%透明质酸钠造成视网膜脱离,对照组小鼠18只左眼仅作巩膜穿刺。分别于手术后1、3、7和28 d摘除眼球,视网膜切片进行组织化学、免疫荧光染色,共聚焦显微镜检查。抗视锥和抗视杆细胞的抗体分别标记视锥和视杆细胞,dUTP缺口末段标记法(TUNEL)标记凋亡细胞。通过计数存活和凋亡的视锥和视杆细胞来定量光感受器细胞的凋亡和细胞丢失。 结果 凋亡细胞只存在于脱离部分视网膜的外核层,凋亡细胞在视网膜脱离后1 d即可检测得到,3 d时达到高峰,7 d后陡然减少。视网膜脱离后视杆和视锥细胞的死亡呈现同样的时程。 结论 凋亡是视网膜脱离后光感受器细胞死亡的主要病理改变。 (中华眼底病杂志, 2006, 22: 124-127)  相似文献   

4.
高玲  姜德咏 《眼科研究》1999,17(1):41-43
探讨视网膜下间隙的免疫学特点,方法DBA/2鼠视网膜加或不加免疫佐剂、人外周血单个核细胞移植到C5YBL/6或BALB/C鼠的视多膜下间隙。2周后行眼组织病理学检查。结果视网的植物存活,无炎性细胞浸润。人PBMC移植发生免疫排斥反应。结论视网膜下间隙移植,发生一定限度的免疫赦免。  相似文献   

5.
王薇  潘峰  沈丽  杨磊 《眼科研究》2001,19(3):226-228
目的 通过纯视细胞移植,观察重建视网膜神经传导通路中兴奋性神经递质谷氨酸的分布和变化。方法取同龄Wistar鼠和皇家外科学院鼠(RCS)为供/受体,准分子激光切削法制备视细胞层,外路途径移入RCS鼠的视网膜下腔,术后两周取RCS鼠手术眼,手术对照眼,RCS对照眼,Wistar鼠眼。分别做冰冻切片,免疫组织化学染色法染色,普通光学显微镜下观察。结果在视细胞移植术后两周,移植视细胞存活,外丛状层重建,与手术对照眼、RCS对照眼相比,受体RCS鼠重建视网膜中在内丛状层和节细胞层Wistar鼠谷氨酸免疫反应阳性的神经纤维相对密度增多。结论视细胞移植不仅可以使RCS鼠视网膜重建正常的解剖结构,而且在移植后的视网膜中观察到兴奋性神经递质谷氨酸的分布接近正常。  相似文献   

6.
大鼠骨髓间充质干细胞视网膜下移植观察   总被引:9,自引:2,他引:7  
目的:鉴定骨髓间充质干细胞(mesenchymal stem cells,MSCs)在体外不诱导的条件下视网膜下移植后的定位。方法:体外培养雄性大鼠MSCs,直接作为细胞供体视网膜下移植于成年雄性大鼠视网膜下,4周后将动物处死,取眼球做石蜡切片,用Y染色体鉴定,为做进一步验证,将MSCs用重组腺相关病毒AAV-gfp感染后移植,分别于4、8周取动物眼球作冰冻切片,于荧光显微镜下做绿色荧光蛋白(green fluorescence protein,GFP)表达观察。结果:培养的MSCs集落生长迅速,均一性好;Y染色体原位杂交鉴定表明来源于MSCs的阳性细胞融合入了原来的视网膜结构,在光学显微镜下可分布于视锥,视杆细胞层,双极细胞层及节细胞层;在荧光显微镜下可见GFP标记的阳性细胞存在,分布于视网膜色素上皮层、视锥,视杆细胞层,双极细胞层及节细胞层,细胞形态与结构同周围的视网膜相似;2种标记方法检测到的视网膜结构完整,未见到玫瑰花结样结构。结论:MSCs可在视网膜下移植后4周与原视网膜结构相融合,2种方法检测到的阳性细胞分布于视网膜色素上皮层,视细胞层,双极细胞层及节细胞层。  相似文献   

7.
潘峰  王薇  张惠蓉  由德勃  任蕾  沈丽 《眼科研究》2000,18(5):412-414
目的 观察正常Wistar大鼠和患遗传性视网膜病变的RCS鼠视网膜内一氧化氮合酶(NOS)的分布。方法 74天Wistar鼠和74天RCS鼠各6眼分为2组,冰冻切片,用NADPH黄递酶组织染色法(NDP),光镜下观察。结果 一氧化氮合酶主要存在于Wistar鼠,RCS鼠的无长突细胞内,在双侧锯齿缘、赤道部和后极部内丛层均有分布,二者间的数量有差异。结论 一氧化氮合酶分布在Wistar鼠,RCS鼠的无长突细胞,但视细胞萎缩对含一氧化氮合酶的阳性无长突细胞数量有影响。  相似文献   

8.
经巩膜外路至视网膜下腔移植视网膜细胞的实验研究   总被引:1,自引:1,他引:0  
Tang SB  Luo Y  Li JQ  Bai NY  Yang B  Wang Z  Zheng HL 《中华眼科杂志》2003,39(10):610-615
探讨视网膜光感受器细胞的移植方法及临床意义。方法将16只昆明鼠随机分为A组和B组,每组均8只鼠。于手术显微镜下,用特殊显微注射器穿过巩膜、脉络膜,在A组昆明鼠的视网膜下腔注入视网膜混合细胞,在B组昆明鼠的视网膜下腔注入纯光感受器细胞。于移植术后30、90及180 d摘除实验眼,于光镜下观察移植细胞在视网膜下腔生长的情况。结果大多数标本(13/15)HE染色显示视网膜细胞准确移植在受体眼的视网膜下腔,未见炎性细胞浸润和受体视网膜破坏;且移植到受体视网膜下腔的细胞在术后180 d仍存活。仅少数(2/15)标本可见受体视网膜结构破坏。移植的视网膜混合细胞均形成“玫瑰花”样结构,而移植的纯视网膜光感受器细胞则在视网膜下腔形成整齐的细胞层。结论经巩膜外路至视网膜下腔的显微注射法是较为理想的视网膜下腔注射给药和视网膜细胞移植方式。纯视网膜光感受器细胞移植后的生长状况和功能接近正常生理状态的视网膜组织结构,为临床治疗视网膜变性疾病提供了新途径。  相似文献   

9.
次全视网膜冷凝联合抗代谢物治疗新生血管性青光眼   总被引:5,自引:2,他引:3  
目的:探讨新生血管性青光眼尤其晚期病例的治疗方法,方法:将40例新生血管性表示我眼随机分为次全视网膜冷凝(SRC)组23例(23只眼),单纯用SRC或联合抗青光眼手术,对照组17例(17只眼)行抗青光眼手术,两组术后均局部应用5-Fu和地塞米松,术后随访1月~5年,平均6个月。结果:SRC组的治疗成功率(73.9%_高于对照组(41.2%),差异有显著性(P〈0.05)。结论:单纯次全视网膜冷凝术  相似文献   

10.
小鼠光性损伤后视网膜神经上皮层移植的初步探讨   总被引:1,自引:0,他引:1  
Peng Q  Ren P  Liu S 《中华眼科杂志》1999,35(4):265-267,I016
目的 探讨光性损伤后视网膜光感受器能否被拯救的可能性。方法 用自制的光损伤箱建立网膜光性损伤的小鼠模型,将光照后的60只小鼠分为2组,Ⅰ组20只为光损伤组,Ⅱ组40只为移植组,又将Ⅱ组分为ⅡA组20只鼠和ⅡB组20只鼠,分别于光性损伤后20h和1周上经路方法行视网膜神经上皮层移植,分别于术后6、12、18及30天时处死受体鼠,用光镜检查移植物是否能救光性损伤后受体鼠视网膜的光感受器。结果 Ⅰ组,可  相似文献   

11.
视网膜视细胞的成片移植   总被引:2,自引:0,他引:2  
目的 探索用准分子激光切削技术制备视网膜单层细胞植片,经内入路视网膜下腔的单层视细胞成片移植。方法 用准分子激光对大鼠视网膜进行切削,制取单层视细胞植片,此后,按内入路手术方法进行了兔视网膜下腔的异种移植。结果 切削后所得视细胞植片由单层视细胞组成,结构完整,包括外丛状层、外核层和外节层;视细胞植片经明胶包埋后被准确植入宿主视网膜下腔中,移植术后第1,2天宿主观视网膜未能复位,呈脱离状态,移植物没能与视网膜色素上皮层相贴;移植后10天,宿主视网膜复位,视细胞移植片平铺于宿主视网膜下腔中,植片视细胞外节也宿主视网膜色素上皮层相贴;移植后10天,宿主视网膜复位,视细胞移植片平铺于宿主视网膜下腔中,植片视细胞外节与宿主视网膜色素上皮层相贴,未见明显免疫排异现象。结论 准分子激光制备单层视细胞植片方法简单、可行;初步观察到内入路单层视细胞成片移植后,视细胞植片能够在宿主视网膜下腔中以正常生理位置存活;视网膜下腔为理想的视网膜移植的受位。  相似文献   

12.
This study investigated whether transplanted sheets of human fetal retina together with its retinal pigment epithelium (RPE) could develop and maintain their cytoarchitecture after long survival times. Transplant recipients were nine albino athymic nu/nu rats with a normal retina. The donor tissue was dissected from fetuses of 12-17 weeks gestational age. Transplants were analyzed at 5-12 months after surgery by light and electron microscopy, and immunohistochemistry with various antibodies specific for rhodopsin, S-antigen, transducin, neurofilament and synaptophysin. In 4 of 11 transplants, the RPE stayed as a monolayer sheet and supported the development of the retinal sheet with a normal lamination, including photoreceptor inner and outer segments. Cones and rods in the organized transplants were labeled with different photoreceptor markers. Inner and outer plexiform layers, containing cone pedicles and rods spherules, were immunoreactive for synaptophysin. As the recipients had a normal retina, transplant/host integration was not expected. However, at the transplant/host interface, there were sometimes areas without glial barriers, and neurofilament-containing processes could be observed crossing between transplant and host. In other, more disorganized transplants, the RPE cells were partially dispersed or clumped together in clusters. Such transplants developed photoreceptors in rosettes, often with inner and outer segments.In conclusion, sheets of human fetal retina transplanted together with its RPE to the subretinal space of nude rats can develop and maintain perfectly laminated transplants after long survival times, indicating the potential of applying cotransplantation to human patients with retinal diseases.  相似文献   

13.
When aggregates of immature retina are transplanted to lesioned adult retina, the donor cells reorganize themselves into folded sheets and rosettes. With the exception of retinal ganglion cells and an inner limiting membrane, most cell types and all layers develop, corresponding to the normal retina. The transplants can integrate with the adult host retina without the presence of glial barriers. Mouse and human donor retinas have been transplanted to immunosuppressed rat hosts with long-term survival. Transplants of embryonic retina cografted with RPE can contain an apparent ganglion cell layer and an inner limiting membrane, which are not observed in transplants of retina alone. Embryonic rat donor retinas can be transplanted successfully after long-term storage in liquid nitrogen.  相似文献   

14.
目的探讨骨髓基质干细胞(bone marrow stromal cell,BMSC)在新生S334转基因视网膜变性大鼠和(Sprague-Dawley)SD大鼠眼内生存、发育、分化的情况.方法人BMSC培养在含10%胎牛血清的(a-Modified Eagle Medium,a-MEM)培养基上扩增;实验分四组;第一组S334转基因鼠,细胞移植联合维甲酸(Retinoid acid)RA注射组;第二组S334转基因大鼠,细胞移植组;第三组SD大鼠,细胞移植联合RA注射组;第四组SD大鼠,细胞移植组.以上每组各5只.2μl细胞混悬液(约4×104个细胞)注入生后1 d大鼠的玻璃体腔.于生后14 d,生后23 d处死动物,取出眼球作塑料切片,进行组织学分析.结果本实验结果表明,BMSC移植到生后1 d S334杂合子转基因大鼠玻璃体腔,在外源性RA的作用下,可能参与宿主视网膜的后期发育,可见内核层细胞增多,整个神经视网膜增厚,但感光细胞层数不变.而无外源性RA移植组,在生后14 d时,可见移植细胞形成类似血岛样结构,而向神经分化的成分少.SD组,在联合RA注射时,生后23 d将动物处死,作组织学分析,发现移植细胞在眼内能移行分化,也可见视网膜内核层细胞增多,同时由于RA的作用,可见感光细胞增生.在无RA作用时,可见宿主神经视网膜结构紊乱,移植细胞增生形成非典型性增生细胞团.结论本实验初步研究结果提示,BMSC移植到新生S334视网膜变性转基因大鼠和SD大鼠眼内,能参与宿主视网膜的发育,并有向视网膜内核层细胞分化的可能性.将为自体细胞移植治疗视网膜变性性疾病提供又一细胞来源.  相似文献   

15.
Photoreceptors from neonatal transgenic mice with normally developing retinas were transplanted to the subretinal spaces of 2-3-month-old rd mutant mice that lack photoreceptors. The transgenic mouse photoreceptors express high levels of the lac Z reporter gene product, beta-galactosidase, which facilitated tracking the transplanted cells. Two sources were used for these cells: (1) dissection of retinal microaggregates containing photoreceptors and (2) papain-dissociated photoreceptors. Host retinas were examined after transplantation. Both methods led to survival of photoreceptors for at least 2 mo after transplantation. Relatively mature outer segments were found only in transplanted microaggregates; this occurred optimally when the cells were adjacent to the retinal pigment epithelium (RPE). beta-galactosidase-labeled outer segments associated closely with the apical processes of the host RPE, which, together with labeled phagosomes in the RPE cells, suggested functional interaction between the transplanted photoreceptors and the host RPE. This study is the first to the authors' knowledge to show electron microscopically that a morphologically normal-appearing photoreceptor layer can be reconstructed in an otherwise photoreceptorless retina.  相似文献   

16.
Retinal pigment epithelial (RPE) cells isolated from 6-8-day-old pigmented Long-Evans rat eyes were successfully grafted onto Bruch's membrane in albino Sprague-Dawley hosts ranging in age from 10 days postnatal to young adulthood. Injections of RPE cells were made into the subretinal space using a lesion paradigm which penetrates through the dorsal surface of the eye cutting through the sclera and choroid. Suspensions of pigmented RPE cells were labeled with Nuclear Yellow prior to transplantation, and at 1 week after grafting, the transplanted RPE cells were found attached to previously denuded areas of Bruch's membrane. The grafted RPE cells were positively identified by double labeling-only those RPE cells with melanosomes in their cytoplasm showed fluorescent Nuclear Yellow labeling; host albino RPE cells showed no nuclear labeling. The grafted RPE cells developed a normal relationship with photoreceptor cell outer segments as seen by electron microscopy. When compared with intact retinas or host control areas there were no significant differences in the thickness of the outer nuclear layer or in the lengths of photoreceptor inner and outer segments underneath the grafted RPE cells for at least 3 months after transplantation, which was the longest survival time examined.  相似文献   

17.
Retinal transplantation--advantages of intact fetal sheets   总被引:2,自引:0,他引:2  
Retinal transplantation aims to prevent blindness and to restore eyesight, i.e., to rescue photoreceptors or to replace damaged photoreceptors with the hope of reestablishing neural circuitry. Retinal donor tissue has been transplanted as dissociated cells or intact sheets. A promising experimental paradigm is the subretinal transplantation of sheets of fetal retina with or without its attached retinal pigment epithelium (RPE) into recipient rats with retinal degeneration. As long as healthy RPE either from the host or from the graft is present, such transplants can develop lamination resembling a normal retina. Different methods have been used to demonstrate transplant/host connectivity. In two different rat retinal degeneration models, visually evoked responses can be demonstrated in an area of the superior colliculus corresponding to the placement of the transplant in the retina. In summary, sheets of fetal retina can morphologically repair an area of a degenerated retina, and there is evidence to suggest that transplants form synaptic connections with the host and restore visual responses in blind rats.  相似文献   

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