Paris-Saclay University, Gustave Roussy Institute, Institut National de la Santé et de la Recherche Médicale Inserm U1170, Villejuif, France.
Department of Biological and Chemical Engineering, National Institute of Applied Sciences and Technology, Tunis, Tunisia.
Methods Mol Biol. 2021;2308:107-115. doi: 10.1007/978-1-0716-1425-9_9.
The intrinsic properties of self-renewal and differentiation of hematopoietic stem and progenitor cells (HSPCs) play a critical role in the regeneration of mature hematopoietic cells at steady state and in stress conditions including bleeding, inflammation and aging. Common techniques such as flow cytometry and genetic methods have answered many questions about their intrinsic and extrinsic regulation. Using these approaches, it was demonstrated that HSPCs in the bone marrow demonstrate low rates of proliferation and apoptosis. This dormant phenotype is associated with a low production of reactive oxygen species and low mitochondrial activity.Here, we describe the methodology to characterize the physiologic state of HSPCs isolated from their native hematopoietic organ using flow cytometry-based assays. These protocols allow evaluation of their ROS levels and activated signaling pathways under various conditions.
造血干细胞和祖细胞（HSPCs）的自我更新和分化的内在特性在稳态下和在包括出血、炎症和衰老在内的应激条件下成熟造血细胞的再生中起着关键作用。流式细胞术和遗传方法等常见技术已经回答了许多关于它们的内在和外在调节的问题。使用这些方法，已经证明骨髓中的 HSPCs 增殖和凋亡的速度较低。这种休眠表型与活性氧（ROS）的产生低和线粒体活性低有关。在这里，我们描述了使用基于流式细胞术的测定法从其天然造血器官分离 HSPCs 来描述其生理状态的方法学。这些方案允许在各种条件下评估它们的 ROS 水平和激活的信号通路。
