La Torre Anna, Lamba Deepak A, Jayabalu Anu, Reh Thomas A
Department of Biological Structure, Institute for Stem Cells and Regenerative Medicine, University of Washington, Seattle, WA, USA.
Methods Mol Biol. 2012;884:229-46. doi: 10.1007/978-1-61779-848-1_16.
Over the last few years, numerous studies have introduced strategies for the generation of neuronal populations from embryonic stem cells. These techniques are valuable both in the study of early neurogenesis and in the generation of an unlimited source of donor cells for replacement therapies. We have developed a protocol to direct mouse and human embryonic stem cells to retinal fates by using the current model of eye specification. Our method is a multistep protocol in which the cultures are treated with IGF1 and a combination of BMP and Wnt inhibitors to promote the expression of key retinal progenitor genes, as assayed by RT-PCR and immunofluorescence microscopy. The retinal progenitor population spontaneously undergoes differentiation towards various types of retinal neurons, including photoreceptors.
在过去几年中，众多研究介绍了从胚胎干细胞生成神经元群体的策略。这些技术在早期神经发生的研究以及为替代疗法生成无限的供体细胞来源方面都很有价值。我们利用当前的眼发育模型，开发了一种将小鼠和人类胚胎干细胞定向诱导为视网膜命运细胞的方案。我们的方法是一个多步骤方案，其中通过RT-PCR和免疫荧光显微镜检测，用IGF1以及BMP和Wnt抑制剂的组合处理培养物，以促进关键视网膜祖细胞基因的表达。视网膜祖细胞群体可自发地向包括光感受器在内的各种类型的视网膜神经元分化。
