Wassélius J, Ghosh F
Department of Ophthalmology, Lund University Hospital, Sweden.
Invest Ophthalmol Vis Sci. 2001 Oct;42(11):2632-8.
To study the survival of adult retinal grafts prepared in a physiologically optimized way.
Twenty-three rabbits received an adult full-thickness rabbit retinal transplant positioned under the host retina, using a vitrectomy technique. The transplants were prepared using a procedure based on a previously described in vitro model used for physiological experiments on the adult retina. Five rabbits received a fragmented graft. All grafts were prelabeled with 4',6-diaminidin-2-phenylindoldihydrochloride (DAPI) to allow identification. The eyes were examined by light and fluorescence microscopy 6 to 174 days after surgery. To assess the amount of cell death in the graft before actual transplantation, in vitro experiments were performed. The extent of cell death in retinas prepared by the optimized protocol was examined and compared with a simpler preparation previously used successfully for embryonic grafts. The amount of cell death in the in vitro experiments was evaluated using a fluorescent green nucleic acid stain that penetrates dying cells.
In 21 of the 23 animals that received full-thickness grafts prepared in an optimized way, the transplant survived. Sixteen grafts, including all four with a 174-day survival time, displayed normal morphology, with all retinal layers preserved. The fragmented grafts survived poorly. The in vitro experiments showed minimal cell death in retinas prepared according to the optimized protocol, whereas control retinas displayed extensive cell death after 5 hours.
The results showed that it is possible to transplant adult retina in the rabbit and that the grafts survive well if they are prepared under physiologically optimized conditions and the integrity of the grafted tissue is kept intact.
研究以生理优化方式制备的成年视网膜移植物的存活情况。
23只兔子接受了成年兔全层视网膜移植,采用玻璃体切割技术将其置于宿主视网膜下方。移植片是根据先前用于成年视网膜生理实验的体外模型所描述的程序制备的。5只兔子接受了碎片状移植物。所有移植物都预先用4',6-二脒基-2-苯基吲哚二盐酸盐(DAPI)标记以便识别。术后6至174天通过光镜和荧光显微镜检查眼睛。为了评估实际移植前移植物中的细胞死亡量,进行了体外实验。检查了通过优化方案制备的视网膜中的细胞死亡程度,并与先前成功用于胚胎移植物的更简单制备方法进行了比较。使用穿透死亡细胞的荧光绿色核酸染料评估体外实验中的细胞死亡量。
在接受以优化方式制备的全层移植物的23只动物中,有21只的移植物存活。16个移植物,包括所有存活174天的4个移植物,显示出正常形态,所有视网膜层均得以保留。碎片状移植物存活不佳。体外实验表明,根据优化方案制备的视网膜中细胞死亡极少,而对照视网膜在5小时后显示出广泛的细胞死亡。
结果表明,在兔子中移植成年视网膜是可行的,并且如果在生理优化条件下制备移植物并保持移植组织的完整性,移植物可以很好地存活。