Isolation of primitive human hematopoietic progenitors on the basis of aldehyde dehydrogenase activity
Isolation of primitive human hematopoietic progenitors on the basis of aldehyde dehydrogenase activity
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DOI：
10.1073/pnas.96.16.9118
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发表时间：
1999-08-03
影响因子：
11.1
通讯作者：
Smith, C
中科院分区：
文献类型：
--
作者：
Storms, RW;Trujillo, AP;Smith, C
由于造血干细胞富含醛脱氢酶（ALDH）活性，我们开发了一种用于ALDH的荧光底物，称为BODIPY氨基乙醛（BAAA），并测试了其分离原始人造血细胞的潜力。具有低正交光散射和明亮荧光强度的细胞群体（SSC（lo）ALDH（br）细胞）可以容易地从用BAAA和多药耐药抑制剂维拉帕米共染色的人脐带血细胞中分离。SSC（lo）ALDH（br）群体耗尽谱系定型细胞，对于CD 34（+）CD 38（lo/-）细胞，纯度为40-90%，并且在短期和长期培养分析中检测到的原始造血祖细胞富集50- 100倍。总之，这些观察结果表明，使用BAAA基于ALDH活性分级分离人造血干细胞是分离原始人造血祖细胞的有效方法。这种技术也可以用于从其他组织中分离干细胞。
Because hematopoietic stem cells are rich in aldehyde dehydrogenase (ALDH) activity, we developed a fluorescent substrate for ALDH, termed BODIPY aminoacetaldehyde (BAAA), and tested its potential for isolating primitive human hematopoietic cells. A population of cells with low orthogonal light scattering and bright fluorescence intensity (SSC(lo)ALDH(br) cells) could be readily fractionated from human umbilical cord blood cells costained with BAAA and the multidrug-resistance inhibitor verapamil, The SSC(lo)ALDH(br) population was depleted of lineage-committed cells, 40-90% pure for CD34(+)CD38(lo/-) cells, and enriched 50- to 100-fold for primitive hematopoietic progenitors detected in short- and long-term culture analyses. Together, these observations indicate that fractionating human hematopoietic: stem cells on the basis of ALDH activity using BAAA is an effective method for isolating primitive human hematopoietic progenitors. This technique may be useful for isolating stem cells from other tissues as well.