Castillo B V, del Cerro M, White R M, Cox C, Wyatt J, Nadiga G, del Cerro C
Department of Neurobiology and Anatomy, University of Rochester Medical Center, New York 14642, USA.
Exp Neurol. 1997 Jul;146(1):1-9. doi: 10.1006/exnr.1997.6534.
This study determines the efficacy of nonfetal human retinal pigment epithelium (RPE) for photoreceptor rescue utilizing the dystrophic RCS rat as an animal model. Eyes from 10- and 49-year-old donors were obtained through the Rochester Eye and Human Parts Bank. The RPE was isolated by enzymatic treatment of the choroid-RPE with 2% dispase for 30 min at 37 degrees C. Mechanically dissociated RPE cells were injected at the superior hemisphere into the subretinal space of dystrophic RCS rats during the fourth postnatal week. Rats receiving vehicle injection served as sham controls. The animals were immunosuppressed with daily cyclosporine injections (10 mg/kg) and sacrificed 30 days posttransplantation for histologic evaluation of the RPE graft and its effect on photoreceptor survival. Transplantation of adult human RPE promoted the survival of photoreceptors in the dystrophic RCS rat. Morphometric analysis of the grafted superior hemisphere demonstrated a threefold increase in photoreceptor cell density (149.2 +/- 50 SD) compared to sham controls (39.7 +/- 31 SD) and the untouched inferior hemisphere (52.8 +/- 28 SD). RPE from the 49-year-old donor was as effective as RPE from the 10-year-old donor in promoting photoreceptor survival. The results of this study in RCS rats suggests that RPE from adult human donors of varied ages is suitable for transplantation and retains the capability to promote survival of photoreceptor cells. This finding opens the possibility of using nonfetal RPE cells in human retinal transplantation.
本研究以营养不良的RCS大鼠作为动物模型，确定非胎儿人类视网膜色素上皮（RPE）对光感受器挽救的疗效。通过罗切斯特眼及人体组织库获取10岁和49岁供体的眼睛。用2%的中性蛋白酶在37℃下对脉络膜-RPE进行酶处理30分钟，分离出RPE。在出生后第四周，将机械解离的RPE细胞注入营养不良的RCS大鼠上半球的视网膜下间隙。接受载体注射的大鼠作为假手术对照。每天给动物注射环孢素（10mg/kg）进行免疫抑制，并在移植后30天处死，用于对RPE移植物及其对光感受器存活的影响进行组织学评估。成人人类RPE移植促进了营养不良的RCS大鼠光感受器的存活。对移植的上半球进行形态计量分析表明，与假手术对照（39.7±31标准差）和未处理的下半球（52.8±28标准差）相比，光感受器细胞密度增加了三倍（149.2±50标准差）。49岁供体的RPE在促进光感受器存活方面与10岁供体的RPE一样有效。这项在RCS大鼠中的研究结果表明，不同年龄成人供体的RPE适合移植，并保留了促进光感受器细胞存活的能力。这一发现开启了在人类视网膜移植中使用非胎儿RPE细胞的可能性。
