经巩膜外路至视网膜下腔移植视网膜细胞的实验研究

Subretinal transplantation of retinal cells by outer approach

目的 探讨视网膜光感受器细胞的移植方法及临床意义。方法 将 1 6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 1 80d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 (1 3 1 5)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 1 80d仍存活。仅少数 (2 1 5)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。

Objective To investigate the approaches and effect of pure retinal photoreceptor cells and mixed retinal cells subretinal transplantation on retinal degenerative diseases. Methods 16 Kunming mice were divided into two groups: group A and group B ( n =8). Through sclera and choroid,mixed retinal cells were injected into the subretinal space of mice eyes of group A and pure retinal photoreceptors were injected into the subretinal space of mice eyes of group B by using a specific micro syringe under operative microscopy. 30, 90 and 180 d after transplantation, experimental eyes were enucleated, stained with HE and analyzed by light microscopy. Results Most transplants (13/15) were exactly in host subretinal space without the infiltration of inflammatory cells and interfere of retina. The infiltration of inflammatory cells and damage of retina were only found in few eyes (2/15). The transplanted retinal cells survived well at 180 days after transplantation. The mixed retinal cells all formed rosette, but the pure retinal photoreceptors formed an orderly layer of photoreceptors. Conclusions This specific technique of subretinal microinjection is an easy, safe and optimal technique to meet the need of subretinal drug administration and retinal cells transplantation for retinal degenerative diseases. It is easier for the transplanted pure retinal photoreceptor cells to establish functional connection with host cells through subretinal microinjection.