Phenotypic Analysis of the Mouse Hematopoietic Hierarchy Using Spectral Cytometry: From Stem Cell Subsets to Early Progenitor Compartments.
Phenotypic Analysis of the Mouse Hematopoietic Hierarchy Using Spectral Cytometry: From Stem Cell Subsets to Early Progenitor Compartments.
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DOI：
10.1002/cyto.a.24041
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发表时间：
2020-10
期刊：
影响因子：
--
通讯作者：
Reynaud D
中科院分区：
文献类型：
--
作者：
Solomon M;DeLay M;Reynaud D
通过流式细胞术进行的表型分析是研究造血系统最常用的主要工具之一。在这里，我们提出了一个复杂的面板设计的光谱流式细胞术，允许在小鼠造血干细胞和祖细胞室的深入分析。所开发的面板包括造血干细胞（HSC）室，多能祖细胞与早期标记的谱系规范和一系列的祖细胞与淋巴，粒-巨噬细胞，巨核细胞和红系谱系承诺。它具有已知的关键标记物的内置冗余，通过分离具有不同功能潜力的子集来破译HSC隔室的精细结构。作为一种资源，我们使用这个面板来提供这些表型定义的造血隔室在动物生命期间的演变的快照视图。我们表明，通过使用光谱流式细胞仪，这个面板是兼容的GFP表达基因报告小鼠在造血系统的分析。我们利用这个工具来确定以前描述的标记物，如CD 150，CD 34，CD 105，CD 41，ECPR和CD 49 b如何定义特定的HSC亚群，并确认转录因子Gfi 1的高表达是最原始的HSC区室的标志。总之，我们的研究结果提供了一个方便的协议，在一个分析中获得更广泛的观点在小鼠模型的造血结构。我们的研究结果也可以作为进一步开发高端面板的基础，利用光谱流式细胞术超越本报告中提出的15-荧光染料面板。
Phenotypic analysis by flow cytometry is one of the most utilized primary tools to study the hematopoietic system. Here, we present a complex panel designed for spectral flow cytometry that allows for the in-depth analysis of the mouse hematopoietic stem and progenitor compartments. The developed panel encompasses the hematopoietic stem cell (HSC) compartment, an array of multipotent progenitors with early marks of lineage specification and a series of progenitors associated with lymphoid, granulo-macrophagic, megakaryocytic and erythroid lineage commitment. It has a built-in redundancy for key markers known to decipher the fine architecture of the HSC compartment by segregating subsets with different functional potential. As a resource, we used this panel to provide a snapshot view of the evolution of these phenotypically defined hematopoietic compartments during the life of the animals. We show that by using a spectral cytometer, this panel is compatible with the analysis of GFP-expressing gene-reporter mice across the hematopoietic system. We leverage this tool to determine how previously described markers such as CD150, CD34, CD105, CD41, ECPR, and CD49b define specific HSC subsets and confirm that high expression of the transcription factor Gfi1 is a hallmark of the most primitive HSC compartment. Altogether, our results provide a convenient protocol to obtain in one analysis a more extensive view of the hematopoietic architecture in mouse models. Our results could also serve as a base for further development of high-end panels leveraging spectral flow cytometry beyond the 15-fluorochrome panel presented in this report.
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影响因子：
82.9
作者：
Gur-Cohen S;Itkin T;Chakrabarty S;Graf C;Kollet O;Ludin A;Golan K;Kalinkovich A;Ledergor G;Wong E;Niemeyer E;Porat Z;Erez A;Sagi I;Esmon CT;Ruf W;Lapidot T
通讯作者：
Lapidot T
影响因子：
64.8
作者：
Rodriguez-Fraticelli AE;Wolock SL;Weinreb CS;Panero R;Patel SH;Jankovic M;Sun J;Calogero RA;Klein AM;Camargo FD
通讯作者：
Camargo FD
影响因子：
64.5
作者：
Kiel, MJ;Yilmaz, ÖH;Morrison, SJ
通讯作者：
Morrison, SJ
DOI：
10.1007/978-1-4939-7346-0_13
发表时间：
2018-01-01
期刊：
FLOW CYTOMETRY PROTOCOLS, 4TH EDITION
影响因子：
--
作者：
Pronk, Cornelis J. H.;Bryder, David
通讯作者：
Bryder, David
影响因子：
64.8
作者：
Olsson, Andre;Venkatasubramanian, Meenakshi;Chaudhri, Viren K.;Aronow, Bruce J.;Salomonis, Nathan;Singh, Harinder;Grimes, H. Leighton
通讯作者：
Grimes, H. Leighton