Abstract：
Blood cell production arises from the activity of hematopoietic stem cells (HSCs), defined by their self-renewal capacity and ability to give rise to all mature blood cell types. The mouse remains one of the most studied species in hematological research, and markers to define and isolate mouse HSCs are well-established. Given the very low frequency of HSCs in the bone marrow, stem cell pre-enrichment by red blood cell lysis and magnetic cell separation is often performed as part of the isolation process to reduce sorting times. Several pre-enrichment strategies are available, differing in their speed, degree of enrichment, final cell yield, and cost. In the current study, we performed a side-by-side comparison and provide a decision tree to help researchers select a pre-enrichment strategy for mouse HSC isolation depending on their downstream application. We then compared different pre-enrichment techniques in combination with metabolomics analysis of HSCs, where speed, yield and temperature during pre-enrichment are crucial factors, and found that the choice of pre-enrichment strategy significantly impacts the number of metabolites detected and levels of individual metabolites in HSCs.
摘要：
血细胞的产生来自造血干细胞（HSC）的活性，由它们的自我更新能力和产生所有成熟血细胞类型的能力来定义。小鼠仍然是血液学研究中研究最多的物种之一，和用于定义和分离小鼠HSC的标记是良好建立的。鉴于骨髓中HSC的频率非常低，通过红细胞裂解和磁性细胞分离的干细胞预富集通常作为分离过程的一部分进行以减少分选时间。有几种富集前策略可用，不同的速度，丰富程度,最终细胞产量和成本。在目前的研究中,我们进行了并排比较,并提供了决策树,以帮助研究人员根据其下游应用选择用于小鼠HSC分离的预富集策略.然后，我们将不同的预富集技术与HSC的代谢组学分析相结合，在那里速度，预富集过程中的产量和温度是关键因素，并且发现预富集策略的选择显着影响HSC中检测到的代谢物的数量和单个代谢物的水平。
