Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA, USA.
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Methods Mol Biol. 2023;2683:39-51. doi: 10.1007/978-1-0716-3287-1_4.
The ability to differentiate pluripotent stem cells and to generate specific cell types is a long-standing goal of regenerative medicine. This can be accomplished by recreating the developmental trajectories using sequential activation of the corresponding signaling pathways, or more recently-by direct programming of cell identities using lineage-specific transcription factors. Notably, to be functional in cell replacement therapies, generation of complex cell types, such as specialized neuronal sub-types of the brain, requires precise induction of molecular profiles and regional specification of the cells. However, the induction of the correct cellular identity and marker gene expression can be hampered by technical challenges, one of which is the robust co-expression of multiple transcription factors that is often required for correct cell identity specification. Here, we describe in detail a method for co-expression of seven transcription factors required for efficient induction of dopaminergic neurons with midbrain characteristics from human embryonic and induced pluripotent stem cells.
将多能干细胞进行分化并生成特定的细胞类型是再生医学的长期目标。这可以通过使用相应的信号通路的顺序激活来实现，或者最近，通过使用谱系特异性转录因子来直接对细胞身份进行编程来实现。值得注意的是，为了在细胞替代疗法中具有功能，生成复杂的细胞类型，例如大脑中的专门神经元亚型，需要精确诱导分子特征并对细胞进行区域特异性。然而，诱导正确的细胞身份和标记基因表达可能会受到技术挑战的阻碍，其中之一是通常需要多个转录因子的强烈共表达，这对于正确的细胞身份特化是必需的。在这里，我们详细描述了一种从人胚胎和诱导多能干细胞中高效诱导具有中脑特征的多巴胺能神经元的方法，该方法需要共表达七种转录因子。
