Flores Joseph, Cepeda Ivan L, Cornfeldt Michael L, O'Kusky John R, Doudet Doris J
Department of Medicine/Neurology and the Pacific Parkinson's Research Centre, University of British Columbia, 2221 Wesbrook Mall, Vancouver, BC, Canada.
J Neuropathol Exp Neurol. 2007 Jul;66(7):585-96. doi: 10.1097/nen.0b013e318093e53a.
Previous studies have demonstrated that the intrastriatal implantation of human retinal pigment epithelial cells attached to gelatin microcarriers (hRPE-GM) ameliorates behavioral deficits in animal models of Parkinson disease. However, there are only sparse data on cell survival in the host. In this study, we characterized a variety of retinal pigment epithelial (RPE)-specific markers in vitro and used these markers to investigate the long-term survival of hRPE-GM implants. Sprague-Dawley rats (n = 22) were unilaterally lesioned with 6-hydroxydopamine (6-OHDA) and implanted with hRPE-GM without immunosuppression. Rats were euthanized at 48 hours, 7 days, 4 weeks, and 5 months postimplant and immunohistochemically processed using the following antibodies: 1) human-specific nuclear mitotic apparatus protein (NuMA-Ab2), 2) epithelial-specific extracellular matrix metalloproteinase inducer (EMMPRIN), 3) RPE cell-specific RPE65, and the inflammation markers 4) glial fibrillary acidic protein and 5) ED1 (rat CD68). Our analysis revealed NuMA-, EMMPRIN-, and RPE65-immunoreactive cells at different times postimplant. The morphologic features of hRPE cell implants (at 48 hours and 5 months) were confirmed by electron microscopy. Furthermore, despite evidence of chronic inflammation at the later time point, there is an appreciable number of surviving hRPE cells. This study suggests that hRPE-GM implants can survive in the absence of immunosuppression and can be potentially used as an alternative for treating Parkinson disease.
先前的研究表明，将附着于明胶微载体的人视网膜色素上皮细胞（hRPE-GM）纹状体内植入可改善帕金森病动物模型的行为缺陷。然而，关于宿主中细胞存活的数据却很少。在本研究中，我们在体外对多种视网膜色素上皮（RPE）特异性标志物进行了表征，并使用这些标志物来研究hRPE-GM植入物的长期存活情况。将22只Sprague-Dawley大鼠单侧用6-羟基多巴胺（6-OHDA）损伤，并在不进行免疫抑制的情况下植入hRPE-GM。在植入后48小时、7天、4周和5个月对大鼠实施安乐死，并使用以下抗体进行免疫组织化学处理：1）人特异性核有丝分裂器蛋白（NuMA-Ab2），2）上皮特异性细胞外基质金属蛋白酶诱导剂（EMMPRIN），3）RPE细胞特异性RPE65，以及炎症标志物4）胶质纤维酸性蛋白和5）ED1（大鼠CD68）。我们的分析显示在植入后的不同时间存在NuMA、EMMPRIN和RPE65免疫反应性细胞。通过电子显微镜确认了hRPE细胞植入物（在48小时和5个月时）的形态特征。此外，尽管在后期时间点有慢性炎症的证据，但仍有相当数量的存活hRPE细胞。本研究表明，hRPE-GM植入物在没有免疫抑制的情况下可以存活，并有可能用作治疗帕金森病的替代方法。
