Department of Psychiatry, Neurobiology, Pharmacology and Biotechnology, University of Pisa, Pisa, Italy.
PLoS One. 2012;7(3):e33338. doi: 10.1371/journal.pone.0033338. Epub 2012 Mar 14.
In vitro generation of photoreceptors from stem cells is of great interest for the development of regenerative medicine approaches for patients affected by retinal degeneration and for high throughput drug screens for these diseases. In this study, we show unprecedented high percentages of rod-fated cells from retinal stem cells of the adult ciliary epithelium. Molecular characterization of rod-like cells demonstrates that they lose ciliary epithelial characteristics but acquire photoreceptor features. Rod maturation was evaluated at two levels: gene expression and electrophysiological functionality. Here we present a strong correlation between phototransduction protein expression and functionality of the cells in vitro. We demonstrate that in vitro generated rod-like cells express cGMP-gated channels that are gated by endogenous cGMP. We also identified voltage-gated channels necessary for rod maturation and viability. This level of analysis for the first time provides evidence that adult retinal stem cells can generate highly homogeneous rod-fated cells.
从干细胞中体外生成光感受器对于开发针对视网膜变性患者的再生医学方法以及针对这些疾病的高通量药物筛选具有重要意义。在这项研究中，我们展示了来自成年睫状上皮的视网膜干细胞的前所未有的高比例的杆状细胞。杆状细胞的分子特征表明，它们失去了睫状上皮的特征，但获得了光感受器的特征。杆状细胞的成熟在两个水平上进行评估：基因表达和电生理功能。在这里，我们展示了光转导蛋白表达与细胞体外功能之间的强相关性。我们证明体外生成的杆状细胞表达由内源性 cGMP 门控的 cGMP 门控通道。我们还确定了杆状细胞成熟和存活所必需的电压门控通道。这种分析水平首次提供了证据，证明成年视网膜干细胞可以产生高度同质的杆状细胞。
