Klassen Henry, Kiilgaard Jens Folke, Zahir Tasneem, Ziaeian Boback, Kirov Ivan, Scherfig Erik, Warfvinge Karin, Young Michael J
Stem Cell Research, Children's Hospital of Orange County, Orange, California, USA.
Stem Cells. 2007 May;25(5):1222-30. doi: 10.1634/stemcells.2006-0541. Epub 2007 Jan 11.
Work in rodents has shown that cultured retinal progenitor cells (RPCs) integrate into the degenerating retina, thus suggesting a potential strategy for treatment of similar degenerative conditions in humans. To demonstrate the relevance of the rodent work to large animals, we derived progenitor cells from the neural retina of the domestic pig and transplanted them to the laser-injured retina of allorecipients. Prior to grafting, immunocytochemical analysis showed that cultured porcine RPCs widely expressed neural cell adhesion molecule, as well as markers consistent with immature neural cells, including nestin, Sox2, and vimentin. Subpopulations expressed the neurodevelopmental markers CD-15, doublecortin, beta-III tubulin, and glial fibrillary acidic protein. Retina-specific markers expressed included the bipolar marker protein kinase Calpha and the photoreceptor-associated markers recoverin and rhodopsin. In addition, reverse transcription-polymerase chain reaction showed expression of the transcription factors Dach1, Hes1, Lhx2, Pax6, Six3, and Six6. Progenitor cells prelabeled with vital dyes survived as allografts in the subretinal space for up to 5 weeks (11 of 12 recipients) without exogenous immune suppression. Grafted cells expressed transducin, recoverin, and rhodopsin in the pig subretinal space, suggestive of differentiation into photoreceptors or, in a few cases, migrated into the neural retina and extended processes, the latter typically showing radial orientation. These results demonstrate that many of the findings seen with rodent RPCs can be duplicated in a large mammal. The pig offers a number of advantages over mice and rats, particularly in terms of functional testing and evaluation of the potential for clinical translation to human subjects. Disclosure of potential conflicts of interest is found at the end of this article.
对啮齿动物的研究表明，培养的视网膜祖细胞（RPCs）可整合到退化的视网膜中，这提示了一种治疗人类类似退行性疾病的潜在策略。为了证明啮齿动物研究结果与大型动物的相关性，我们从家猪的神经视网膜中提取祖细胞，并将其移植到同种异体受体的激光损伤视网膜中。在移植前，免疫细胞化学分析显示，培养的猪RPCs广泛表达神经细胞粘附分子，以及与未成熟神经细胞一致的标志物，包括巢蛋白、Sox2和波形蛋白。亚群表达神经发育标志物CD-15、双皮质素、β-III微管蛋白和胶质纤维酸性蛋白。表达的视网膜特异性标志物包括双极标志物蛋白激酶Cα以及与光感受器相关的标志物恢复蛋白和视紫红质。此外，逆转录-聚合酶链反应显示转录因子Dach1、Hes1、Lhx2、Pax6、Six3和Six6的表达。预先用活性染料标记的祖细胞作为同种异体移植物在视网膜下间隙存活长达5周（12只受体中有11只），无需外源性免疫抑制。移植的细胞在猪视网膜下间隙表达转导素、恢复蛋白和视紫红质，提示分化为光感受器，或者在少数情况下，迁移到神经视网膜并延伸出突起，后者通常呈放射状排列。这些结果表明，在啮齿动物RPCs中观察到的许多发现可以在大型哺乳动物中重现。猪相对于小鼠和大鼠具有许多优势，特别是在功能测试和临床转化为人类受试者的潜力评估方面。潜在利益冲突的披露见本文末尾。
