Immunophenotypic Identification of Early Myeloerythroid Development
Immunophenotypic Identification of Early Myeloerythroid Development
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DOI：
10.1007/978-1-4939-7346-0_13
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Bryder, David
中科院分区：
文献类型：
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作者：
Pronk, Cornelis J. H.;Bryder, David
来源于多能造血干细胞的髓红细胞限制性前体细胞产生粒细胞、单核细胞、红细胞和/或血小板谱系的成熟细胞。从造血干细胞到成熟后代的发育阶段的高分辨率分析对于能够研究和理解指导各种细胞命运决定的潜在机制非常重要。此外，这种方法为更深入地了解致病事件，如白血病，最常见的特征是在定义的未成熟前体水平停止分化的疾病。在这一章中，我们提供协议和讨论有关的分析和纯化未成熟的骨髓红细胞系的方法，多参数流式细胞术。大量的文献已经证明了类似方法也适用于人类系统的可行性。然而，在这一章中，我们专注于来自C57 BL/6小鼠，其中基于流式细胞术的免疫表型应用程序已得到最广泛的发展骨髓细胞的鉴定。这也应该允许其应用于在此背景下的转基因模型。为了获得最大的重现性，已使用来自商业供应商的试剂建立了所述的所有方案，以在具有工厂标准配置的流式细胞仪上进行分析。
Myeloerythroid-restricted precursor cells, derived from multipotent hematopoietic stem cells, give rise to mature cells of the granulocyte, monocyte, erythroid, and/or thrombocytic lineages. High-resolution profiling of the developmental stages, from hematopoietic stem cells to mature progeny, is important to be able to study and understand the underlying mechanisms that guide various cell fate decisions. Also, this approach opens for greater insights into pathogenic events such as leukemia, diseases that are most often characterized by halted differentiation at defined immature precursor levels. In this chapter, we provide protocols and discuss approaches concerning the analysis and purification of immature myeloerythroid lineages by multiparameter flow cytometry. A wealth of literature has demonstrated the feasibility of similar approaches also for the human system. However, in this chapter, we focus on the identification of bone marrow cells derived from C57BL/6 mice, in which flow cytometry-based immunophenotypic applications have been most widely developed. This should allow also for its application in genetically modified models on this background. For maximal reproducibility, all protocols described have been established using reagents from commercial vendors to be analyzed on a flow cytometer with factory standard configuration.