Generation of retinal pigmented epithelium from iPSCs derived from the conjunctiva of donors with and without age related macular degeneration.
Generation of retinal pigmented epithelium from iPSCs derived from the conjunctiva of donors with and without age related macular degeneration.
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DOI：
10.1371/journal.pone.0173575
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Dutton JR
中科院分区：
文献类型：
--
作者：
Geng Z;Walsh PJ;Truong V;Hill C;Ebeling M;Kapphahn RJ;Montezuma SR;Yuan C;Roehrich H;Ferrington DA;Dutton JR
多能干细胞分化方案的保真度对于来自胚胎和诱导的多能干细胞的细胞的治疗和商业使用是必要的。干细胞技术的最新进展，特别是一系列化学定义的培养液、底物和分化成分的广泛可用，现在允许设计和实施旨在跨多个研究和制造地点复制的完全定义的派生和分化方案。在这份报告中，我们介绍了这些标准在从患有和不患有年龄相关性黄斑变性的捐赠者的结膜来源的IPSCs中产生视网膜色素上皮的应用。来自70-85岁的人类捐赠者的原代结膜细胞被重新编程以获得多个IPSC系，并使用快速和明确的分化方案将其分化为功能性RPE。将已定义的IPSC来源和培养与已定义的RPE分化方案相结合，可重复生成来自每个供体的功能性RPE，而无需针对每个个体进行方案调整。这种标准化的IPSC派生和RPE分化过程的成功验证展示了一种实用的方法，适用于需要从多个个体经济高效地生成RPE的应用，例如药物测试、人口研究或需要患者特定RPE派生的治疗。此外，结膜细胞被确认为从老年人获得IPSCs的体细胞的实际来源。
Fidelity in pluripotent stem cell differentiation protocols is necessary for the therapeutic and commercial use of cells derived from embryonic and induced pluripotent stem cells. Recent advances in stem cell technology, especially the widespread availability of a range of chemically defined media, substrates and differentiation components, now allow the design and implementation of fully defined derivation and differentiation protocols intended for replication across multiple research and manufacturing locations. In this report we present an application of these criteria to the generation of retinal pigmented epithelium from iPSCs derived from the conjunctiva of donors with and without age related macular degeneration. Primary conjunctival cells from human donors aged 70–85 years were reprogrammed to derive multiple iPSC lines that were differentiated into functional RPE using a rapid and defined differentiation protocol. The combination of defined iPSC derivation and culture with a defined RPE differentiation protocol, reproducibly generated functional RPE from each donor without requiring protocol adjustments for each individual. This successful validation of a standardized, iPSC derivation and RPE differentiation process demonstrates a practical approach for applications requiring the cost-effective generation of RPE from multiple individuals such as drug testing, population studies or for therapies requiring patient-specific RPE derivations. In addition, conjunctival cells are identified as a practical source of somatic cells for deriving iPSCs from elderly individuals.