Ex vivo expansion of human mobilized peripheral blood stem cells using epigenetic modifiers.
Ex vivo expansion of human mobilized peripheral blood stem cells using epigenetic modifiers.
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DOI：
10.1111/trf.12904
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发表时间：
2015-04
期刊：
影响因子：
2.9
通讯作者：
Mahmud N
中科院分区：
文献类型：
--
作者：
Saraf S;Araki H;Petro B;Park Y;Taioli S;Yoshinaga KG;Koca E;Rondelli D;Mahmud N
表观遗传修饰可能控制造血干细胞（HSC）的命运。染色质修饰剂（CMA）、5-氮杂-2 '-脱氧胞苷（5azaD）和曲马斯他丁A（TSA）先前已显示出从脐带血和骨髓扩增HSC。在这里，我们评估CMA是否也可以扩增存在于生长因子动员的人外周血（MPB）中的HSC。在细胞因子的存在下，将5azaD和TSA依次加入到CD 34 + MPB细胞中，并将细胞培养9天。培养后，在5azaD/TSA扩增的细胞中观察到CD 34 + CD 90+细胞扩增3.6 ± 0.5倍，原始集落形成单位（CFU）混合物扩增10.1 ± 0.5倍，长期鹅卵石区形成细胞（CAFC）扩增2.2 ± 0.5倍。相比之下，在没有5azaD/TSA的细胞因子中培养的细胞没有显示扩增;相反，观察到CD 34 + CD 90+细胞（0.7 ± 0.1倍）和CAFC（0.3 ± 0.1）从其初始数量减少。与对照组相比，在5azaD/TSA扩增的MPB细胞中观察到与HSC自我更新相关的几个基因（包括HOXB 4、GATA 2和EZH 2）的转录水平增加相对应的整体低甲基化。5azaD/TSA扩增的MPB细胞保留了体内造血移植能力。可以使用5azaD/TSA扩增来自供体的MPB CD 34+细胞，并且这些扩增的细胞保留体内造血重建能力。这一策略可能被证明是潜在的有用的增加HSC的数量为患者谁不能动员。
Epigenetic modifications likely control fate of hematopoietic stem cells (HSC). The chromatin modifying agents (CMA), 5-aza-2’-deoxyctidine (5azaD) and trichostatin A (TSA) have previously been shown to expand HSC from cord blood and bone marrow. Here we assessed whether CMA can also expand HSCs present in growth factor mobilized human peripheral blood (MPB). 5azaD and TSA were sequentially added to CD34+ MPB cells in the presence of cytokines and the cells were cultured for nine days. Following culture, a 3.6 ± 0.5 fold expansion of CD34+CD90+ cells, a 10.1 ± 0.5 fold expansion of primitive colony forming unit (CFU)-mix, and a 2.2 ± 0.5 fold expansion of long-term cobble stone-area forming cells (CAFC) was observed in 5azaD/TSA expanded cells. By contrast, cells cultured in cytokines without 5azaD/TSA displayed no expansion; rather a reduction in CD34+CD90+ cells (0.7 ± 0.1 fold) and CAFCs (0.3 ± 0.1) from their initial numbers was observed. Global hypomethylation corresponding with increased transcript levels of several genes implicated in HSC self-renewal, including HOXB4, GATA2, and EZH2, was observed in 5azaD/TSA expanded MPB cells in contrast to controls. 5azaD/TSA expanded MPB cells retained in vivo hematopoietic engraftment capacity. MPB CD34+ cells from donors can be expanded using 5azaD/TSA and these expanded cells retain in vivo hematopoietic reconstitution capacity. This strategy may prove to be potentially useful to augment HSCs numbers for patients who fail to mobilize.
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