ASSESSMENT OF ALDEHYDE DEHYDROGENASE IN VIABLE CELLS
ASSESSMENT OF ALDEHYDE DEHYDROGENASE IN VIABLE CELLS
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DOI：
10.1182/blood.v85.10.2742.bloodjournal85102742
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发表时间：
1995-05-15
期刊：
影响因子：
20.3
通讯作者：
HILTON, J
中科院分区：
文献类型：
--
作者：
JONES, RJ;BARBER, JP;HILTON, J
胞质醛脱氢酶（ALDH）是一种负责氧化细胞内醛的酶，在乙醇、维生素A和环磷酰胺代谢中具有重要作用。这种酶在来自多种组织（包括骨髓和肠道）的原始干细胞中高表达，似乎是这些细胞对环磷酰胺产生耐药性的重要机制。然而，尽管造血干细胞 (HSC) 表达高水平的胞质 ALDH，但由于 ALDH 是一种细胞内蛋白，因此通过其 ALDH 表达分离活的 HSC 是不可能的。我们发现荧光醛，丹酰氨基乙醛 (DAAA)，可用于流式细胞术实验，根据 ALDH 含量分离活的小鼠和人类细胞。与 DAAA 孵育后，细胞表现出的丹酰荧光水平与通过蛋白质印迹测定的胞质 ALDH 水平以及细胞对环磷酰胺的敏感性平行。此外，DAAA 似乎是比蛋白质印迹法更灵敏的评估胞质 ALDH 水平的方法。用 DAAA 治疗的骨髓祖细胞增殖正常。此外，与 DAAA 孵育后表达高水平丹磺酰荧光的骨髓细胞富集了造血祖细胞。分离表达高水平胞质 ALDH 的活细胞的能力可能是鉴定和纯化 HSC 以及研究环磷酰胺耐药肿瘤细胞群方法的重要组成部分。 (C) 1995 年，美国血液学会。
Cytosolic aldehyde dehydrogenase (ALDH), an enzyme responsible for oxidizing intracellular aldehydes, has an important role in ethanol, vitamin A, and cyclophosphamide metabolism. High expression of this enzyme in primitive stem cells from multiple tissues, including bone marrow and intestine, appears to be an important mechanism by which these cells are resistant to cyclophosphamide. However, although hematopoietic stem cells (HSC) express high levels of cytosolic ALDH, isolating viable HSC by their ALDH expression has not been possible because ALDH is an intracellular protein. We found that a fluorescent aldehyde, dansyl aminoacetaldehyde (DAAA), could be used in flow cytometry experiments to isolate viable mouse and human cells based on their ALDH content. The level of dansyl fluores-cence exhibited by cells after incubation with DAAA paralleled cytosolic ALDH levels determined by Western blotting and the sensitivity of the cells to cyclophosphamide. Moreover, DAAA appeared to be a more sensitive means of assessing cytosolic ALDH levels than Western blotting. Bone marrow progenitors treated with DAAA proliferated normally. Furthermore, marrow cells expressing high levels of dansyl fluorescence after incubation with DAAA were enriched for hematopoietic progenitors. The ability to isolate viable cells that express high levels of cytosolic ALDH could be an important component of methodology for identifying and purifying HSC and for studying cyclophosphamide-resistant tumor cell populations. (C) 1995 by The American Society of Hematology.