Department of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa Brain and Mind Research Institute, University of Ottawa, Ottawa, ON, Canada.
Methods Mol Biol. 2022;2515:117-127. doi: 10.1007/978-1-0716-2409-8_8.
Adult neural stem and progenitor cells reside in the neurogenic niche of the adult brain and have tremendous potential in regenerative medicine. Compelling evidence suggests that adult neurogenesis plays an important role in hippocampal memory formation, plasticity, and mood regulation. Understanding the mechanisms that regulate the function of neural stem/progenitor cells within the brain is a critical step for the development of regenerative strategies to maintain or enhance neurological function. A major challenge in studying these cells is the limited cell number of adult neural stem cells, and the significant changes in their properties induced by in vitro culture and expansion. To best understand the regulation of these cells, they must be studied within their niche context. In this chapter, we provide a simplified protocol for the harvest and isolation of neural stem cell lineages directly from the murine brain, to provide input material for single-cell RNA-seq. This approach will elucidate the true transcriptional signatures and activated pathways in neural stem cell lineages, within the context of their niche environment.
成年神经干细胞和祖细胞存在于成年大脑的神经发生龛中，在再生医学中有巨大的潜力。令人信服的证据表明，成年神经发生在海马体记忆形成、可塑性和情绪调节中发挥重要作用。了解调节大脑内神经干细胞/祖细胞功能的机制是开发维持或增强神经功能的再生策略的关键步骤。研究这些细胞的主要挑战是成年神经干细胞的细胞数量有限，以及体外培养和扩增引起的其特性的显著变化。为了更好地理解这些细胞的调节，必须在其龛位环境中对其进行研究。在本章中，我们提供了一个简化的方案，用于直接从鼠脑中收获和分离神经干细胞谱系，为单细胞 RNA-seq 提供输入材料。这种方法将阐明神经干细胞谱系在其龛位环境中的真实转录特征和激活途径。
