Hypoxia Enhances the Generation of Retinal Progenitor Cells from Human Induced Pluripotent and Embryonic Stem Cells
Hypoxia Enhances the Generation of Retinal Progenitor Cells from Human Induced Pluripotent and Embryonic Stem Cells
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DOI：
10.1089/scd.2011.0225
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发表时间：
2012-05-01
影响因子：
4
通讯作者：
Veraitch, Farlan S.
中科院分区：
文献类型：
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作者：
Bae, Daekyeong;Mondragon-Teran, Paul;Veraitch, Farlan S.
从人类多能干细胞中有效分化视网膜细胞仍然是开发成功且具有成本效益的各种形式失明细胞疗法的主要挑战。目前的分化策略依赖于将多能干细胞暴露于可溶性生长因子，这些生长因子在20%的氧气（O-2）下在早期发育期间发挥关键作用（如DKK-1、头蛋白和IGF-1）。然而，这种O-2张力在器官发生过程中明显高于O-2水平，并可能损害分化过程。在这项研究中，我们研究了模拟生理O-2张力（2%）对人诱导多能干细胞（iPSC）和人胚胎干细胞（hESC）生成视网膜祖细胞（RPC）的影响。两种细胞类型在20%和2%O-2下诱导分化为RPC。在悬浮培养3天后，作为胚状体（EBs），2%O-2引起缺氧诱导因子响应基因VEGF和LDHA的激活，并伴随着早期眼场基因Six 3和Lhx 2的表达水平升高。在贴壁培养中接种EB后21天，我们观察到在2%O-2下共表达Pax 6和Chx 10的RPC更多。定量聚合酶链反应分析证实，与20%O-2相比，降低O-2张力引起了两种基因表达的上升。我们的研究结果表明，模拟生理O-2是从hiPSC和hESC有效生成RPC的有利条件。
The efficient differentiation of retinal cells from human pluripotent stem cells remains a major challenge for the development of successful and cost-effective cellular therapies for various forms of blindness. Current differentiation strategies rely on exposing pluripotent stem cells to soluble growth factors that play key roles during early development (such as DKK-1, Noggin, and IGF-1) at 20% oxygen (O-2). This O-2 tension is, however, considerably higher than O-2 levels during organogenesis and may impair the differentiation process. In this study, we examined the effect of mimicking the physiological O-2 tension (2%) on the generation of retinal progenitor cells (RPCs) from human induced pluripotent stem cells (iPSCs) and human embryonic stem cells (hESCs). Both cell types were induced to differentiate into RPCs at 20% and 2% O-2. After 3 days in suspension culture as embryoid bodies (EBs), 2% O-2 caused the activation of hypoxia inducible factor responsive genes VEGF and LDHA and was accompanied by elevated expression levels of the early eye field genes Six3 and Lhx2. Twenty-one days after plating EBs in an adherent culture, we observed more RPCs co-expressing Pax6 and Chx10 at 2% O-2. Quantitative polymerase chain reaction analysis confirmed that lowering O-2 tension had caused a rise in the expression of both genes compared with 20% O-2. Our results indicate that mimicking physiological O-2 is a favorable condition for the efficient generation of RPCs from both hiPSCs and hESCs.