Jeffrey Cheah Biomedical Centre, Department of Surgery, Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK.
Institute of Ophthalmology, University College London, London, UK.
Methods Mol Biol. 2022;2520:81-100. doi: 10.1007/7651_2021_401.
Long-term neuroepithelial-like stem cells (lt-NES) derived from human embryonic stem cells are a stable self-renewing progenitor population with high neurogenic potential and phenotypic plasticity. Lt-NES are amenable to regional patterning toward neurons and glia subtypes and thus represent a valuable source of cells for many biomedical applications. For use in regenerative medicine and cell therapy, lt-NES and their progeny require derivation with high-quality culture conditions suitable for clinical use. In this chapter, we describe a robust method to derive multipotent and expandable lt-NES based on good manufacturing practice and cell therapy-grade reagents. We further describe fully defined protocols to terminally differentiate lt-NES toward GABA-ergic, dopaminergic, and motor neurons.
人胚胎干细胞来源的长期神经上皮样干细胞(lt-NES)是一种具有高神经发生潜力和表型可塑性的稳定自我更新祖细胞群体。lt-NES 易于向神经元和神经胶质亚型进行区域性模式化,因此代表了许多生物医学应用中细胞的宝贵来源。为了用于再生医学和细胞治疗,lt-NES 及其后代需要在适合临床应用的高质量培养条件下进行衍生。在本章中,我们描述了一种基于良好生产规范和细胞治疗级试剂的强大方法,用于衍生多能和可扩增的 lt-NES。我们进一步描述了将 lt-NES 终末分化为 GABA 能、多巴胺能和运动神经元的完全定义方案。