Transplantation of Photoreceptor Precursors Isolated via a Cell Surface Biomarker Panel From Embryonic Stem Cell-Derived Self-Forming Retina.
Transplantation of Photoreceptor Precursors Isolated via a Cell Surface Biomarker Panel From Embryonic Stem Cell-Derived Self-Forming Retina.
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DOI：
10.1002/stem.2051
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发表时间：
2015-08
期刊：
影响因子：
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通讯作者：
Sowden JC
中科院分区：
文献类型：
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作者：
Lakowski J;Gonzalez-Cordero A;West EL;Han YT;Welby E;Naeem A;Blackford SJ;Bainbridge JW;Pearson RA;Ali RR;Sowden JC
由于视网膜变性导致的光感受器丧失是无法治愈的失明的主要原因。使用多能干细胞衍生的感光细胞的细胞替代疗法可能是未来可行的治疗方法。实现安全有效的细胞替代关键取决于严格选择和纯化用于移植的最佳细胞。以前，我们证明了荧光报告基因标记的有丝分裂后感光前体细胞的有效移植。由于遗传标记的细胞不适合治疗，因此我们开发了一种表面生物标志物细胞选择策略，用于复杂的多能干细胞分化培养。我们发现，一个五细胞表面生物标志物面板CD 73（+）CD 24（+）CD 133（+）CD 47（+）CD 15（−）促进了从小鼠胚胎干细胞分化的三维自形成视网膜中分离感光细胞前体。重要的是，使用生物标志物组分离的干细胞衍生细胞在视网膜下移植后成功整合并成熟为成年小鼠视网膜中的新视杆细胞。相反，未分选或负选择的细胞在移植后不产生新整合的视杆细胞。生物标志物组还在移植前去除有害的增殖细胞。值得注意的是，我们证明了生物标志物组的表达在人视网膜中是如何保守的，并提出类似的选择策略将有助于分离用于治疗应用的人移植感受态细胞。干细胞2015;33：2469-2482
Loss of photoreceptors due to retinal degeneration is a major cause of untreatable blindness. Cell replacement therapy, using pluripotent stem cell‐derived photoreceptor cells, may be a feasible future treatment. Achieving safe and effective cell replacement is critically dependent on the stringent selection and purification of optimal cells for transplantation. Previously, we demonstrated effective transplantation of post‐mitotic photoreceptor precursor cells labelled by fluorescent reporter genes. As genetically labelled cells are not desirable for therapy, here we developed a surface biomarker cell selection strategy for application to complex pluripotent stem cell differentiation cultures. We show that a five cell surface biomarker panel CD73(+)CD24(+)CD133(+)CD47(+)CD15(−) facilitates the isolation of photoreceptor precursors from three‐dimensional self‐forming retina differentiated from mouse embryonic stem cells. Importantly, stem cell‐derived cells isolated using the biomarker panel successfully integrate and mature into new rod photoreceptors in the adult mouse retinae after subretinal transplantation. Conversely, unsorted or negatively selected cells do not give rise to newly integrated rods after transplantation. The biomarker panel also removes detrimental proliferating cells prior to transplantation. Notably, we demonstrate how expression of the biomarker panel is conserved in the human retina and propose that a similar selection strategy will facilitate isolation of human transplantation‐competent cells for therapeutic application. Stem Cells 2015;33:2469—2482