Improved purification of hematopoietic stem cells based on their elevated aldehyde dehydrogenase activity
Improved purification of hematopoietic stem cells based on their elevated aldehyde dehydrogenase activity
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DOI：
10.3324/haematol.11366
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发表时间：
2007-09-01
期刊：
影响因子：
10.1
通讯作者：
Eaves, Connie
中科院分区：
文献类型：
--
作者：
Christ, Oliver;Lucke, Kai;Eaves, Connie
背景与目的原始人造血细胞比其终末分化的子代细胞具有更高水平的醛脱氢酶（aldehyde dehydrogenase，ALDH）活性，但ALDH水平变化的具体阶段尚未明确。本研究的目的是比较造血细胞分化最早阶段的ALDH水平，并确定是否可以利用这些来获得具有体内长期淋巴-骨髓重建活性的人脐带血细胞的纯度提高。设计和方法首先分析了ALDEFLUOR染色的显示不同ALDH活性水平的人脐带血细胞的各种表面标志物的共表达。这些细胞的亚群，然后分离出多参数流式细胞术和评估短期和长期repopulating活动在亚致死照射immunodeficients.ResultsMost短期骨髓repopulating细胞（STRC-M）和所有长期淋巴骨髓repopulating细胞（LTRC-ML）染色选择性ALDH（+）。STRC-M和LTRC-ML频率的有限稀释分析表明，它们在10%的最高ALDH（+）细胞中富集程度相似且最高。去除表达CD 2、CD 3、CD 7、CD 14、CD 16、CD 24、CD 36、CD 38、CD 56、CD 66 b、或血型糖蛋白A从人脐带血细胞的ALDH低密度级分中分离，具有低的光侧向散射特性，产生了含有LTRC-ML的群体，频率为1/解释和结论ALDH活性升高是原始人脐带血细胞的一个广泛的特性，与其他标志物结合，允许以前所未有的纯度容易地分离干细胞级分。
Background and ObjectivesPrimitive human hematopoietic cells contain higher levels of aldehyde dehydrogenase (ALDH) activity than their terminally differentiating progeny but the particular stages at which ALDH levels change have not been well defined. The objective of this study was to compare ALDH levels among the earliest stages of hematopoietic cell differentiation and to determine whether these could be exploited to obtain improved purity of human cord blood cells with long-term lympho-myeloid repopulating activity in vivo.Design and MethodsALDEFLUOR-stained human cord blood cells displaying different levels of ALDH activity were first analyzed for co-expression of various surface markers. Subsets of these cells were then isolated by multi-parametric flow cytometry and assessed for shortand long-term repopulating activity in sublethally irradiated immunodeficient mice.ResultsMost short-term myeloid repopulating cells (STRC-M) and all long-term lympho-myeloid repopulating cells (LTRC-ML) stained selectively as ALDH(+). Limiting dilution analysis of the frequencies of both STRC-M and LTRC-ML showed that they were similarly and most highly enriched in the 10% top ALDH(+) cells. Removal of cells expressing CD2, CD3, CD7, CD14, CD16, CD24, CD36, CD38, CD56, CD66b, or glycophorin A from the ALDH' low-density fraction of human cord blood cells with low light side-scattering properties yielded a population containing LTRC-ML at a frequency of 1/360.Interpretation and ConclusionElevated ALDH activity is a broadly inclusive property of primitive human cord blood cells that, in combination with other markers, allows easy isolation of the stem cell fraction at unprecedented purities.