LT-HSC Methylcellulose Assay
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Author:
Date:
2014-03-05
[Abstract] Hematopoietic differentiation is a highly complex process originating from an extraordinary population of cells called long-term repopulating hematopoietic stem cells (LT-HSCs). The unique feature of all stem cells, including HSCs, is their exceptional ability to divide asymmetrically giving rise to two different kinds of offspring. One daughter cell becomes an LT-HSC itself (self-renews) to maintain the LT-HSC pool, whereas the second daughter cell pursues a differentiation fate to ultimately give rise to terminally differentiated mature blood cells (Orkin and Zon, 2008). Quantification of phenotypic LT-HSCs can be performed by multi-color flow cytometry and the gold standard for assessment of LT-HSC self-renewal and function is competitive bone marrow transplantation (Miller et al. ...
[摘要] 造血分化是一种非常复杂的过程,源自称为长期重建造血干细胞(LT-HSCs)的非凡细胞群。所有干细胞(包括HSC)的独特之处在于其不对称的划分能够产生两种不同种类的后代。一个子细胞成为LT-HSC本身(自我更新)以维持LT-HSC库,而第二个子细胞追求分化命运,最终产生终末分化的成熟血细胞(Orkin和Zon,2008)。表型LT-HSC的定量可以通过多色流式细胞术进行,用于评估LT-HSC自我更新和功能的黄金标准是竞争性骨髓移植(Miller等,2008)。尽管这些方法对于确定LT-HSC丰度和功能是不可替代的,但它们具有其缺点和局限性。例如,竞争性骨髓移植通常被监测为12-16周内外周血献血者贡献的函数。虽然减少的外周血供体贡献本身表示干/祖细胞隔室中的损伤,但是它不能明确区分降低的LT-HSC自我更新,受损的LT-HSC分化或受损的祖细胞分化。这里我们描述一种LT-HSCs甲基纤维素集落形成测定法,作为直接评估LT-HSC分化能力的快速互补体外方法。如Kerenyi等人所述(2013),该技术是区分LT-HSC或祖细胞分化缺陷的有力工具。
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Mouse Embryonic Stem Cell Maintenance for Differentiation
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Author:
Date:
2011-10-20
[Abstract] Embryonic stem cells are derived from inner cell mass of an embryo that can differentiate into every cell type in the body. Clinically, cultured red blood cell supply is of great interest. However, some of the hurdles need to be overcome. This protocol describes a protocol to maintain mouse stem cells that will be used for hematopoietic lineage differentiation assay in the lab.
[摘要] 胚胎干细胞衍生自胚胎的内细胞团,其可以分化为体内的每种细胞类型。 临床上,培养的红细胞供应是非常有趣的。 然而,一些障碍需要克服。 该协议描述了用于维持小鼠干细胞的方案,其将用于实验室中的造血谱系分化测定。
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