Center for Regenerative Therapies, Technical University Dresden, Dresden, Germany.
PLoS One. 2013;8(1):e54552. doi: 10.1371/journal.pone.0054552. Epub 2013 Jan 24.
A goal in human embryonic stem cell (hESC) research is the faithful differentiation to given cell types such as neural lineages. During embryonic development, a basement membrane surrounds the neural plate that forms a tight, apico-basolaterally polarized epithelium before closing to form a neural tube with a single lumen. Here we show that the three-dimensional epithelial cyst culture of hESCs in Matrigel combined with neural induction results in a quantitative conversion into neuroepithelial cysts containing a single lumen. Cells attain a defined neuroepithelial identity by 5 days. The neuroepithelial cysts naturally generate retinal epithelium, in part due to IGF-1/insulin signaling. We demonstrate the utility of this epithelial culture approach by achieving a quantitative production of retinal pigment epithelial (RPE) cells from hESCs within 30 days. Direct transplantation of this RPE into a rat model of retinal degeneration without any selection or expansion of the cells results in the formation of a donor-derived RPE monolayer that rescues photoreceptor cells. The cyst method for neuroepithelial differentiation of pluripotent stem cells is not only of importance for RPE generation but will also be relevant to the production of other neuronal cell types and for reconstituting complex patterning events from three-dimensional neuroepithelia.
人胚胎干细胞（hESC）研究的目标是将其定向分化为特定的细胞类型，如神经谱系。在胚胎发育过程中，基膜围绕神经板形成紧密的、顶端-基底极性的上皮细胞，然后关闭形成具有单个腔的神经管。在这里，我们展示了在 Matrigel 中三维上皮囊肿培养与神经诱导相结合，可定量转化为含有单个腔的神经上皮囊肿。细胞在 5 天内获得明确的神经上皮特征。神经上皮囊肿自然产生视网膜上皮，部分原因是 IGF-1/胰岛素信号。我们通过在 30 天内从 hESC 中定量产生视网膜色素上皮（RPE）细胞，证明了这种上皮培养方法的实用性。将这种 RPE 直接移植到视网膜变性的大鼠模型中，无需对细胞进行任何选择或扩增，就可以形成由供体衍生的 RPE 单层，挽救感光细胞。多能干细胞神经上皮分化的囊肿方法不仅对 RPE 的生成很重要，而且对其他神经元细胞类型的生成以及对从三维神经上皮重建复杂的模式形成事件也很重要。
