Isolation of a highly quiescent subpopulation of primitive leukemic cells in chronic myeloid leukemia
Isolation of a highly quiescent subpopulation of primitive leukemic cells in chronic myeloid leukemia
复制标题
DOI：
10.1182/blood.v94.6.2056
复制
发表时间：
1999-09-15
期刊：
影响因子：
20.3
通讯作者：
Eaves, A
中科院分区：
文献类型：
--
作者：
Holyoake, T;Jiang, XY;Eaves, A
慢性髓样白血病（CML）的特征是白血病祖细胞的增殖活性增加，产生数量升高的成熟粒细胞。然而，即使是非常高剂量的细胞周期活性剂也无法消除白血病克隆，这表明存在罕见的静态白血病干细胞的子集。要分离此类细胞，我们首先使用Hoechst 33342和吡咯蛋白y染色以获得可行的G（0）和G（1）/S/G（2）/M的CD34（+）细胞的分数，通过荧光激活的细胞分类（FACS） ）从6个慢性相CML患者的样本中，通过证明KI-67和D cyclin表达。在直接分离的GO细胞中鉴定出具有体外祖细胞活性并能够染上免疫缺陷小鼠的白血病（pH+/BCR-ABL（+））细胞。单细胞逆转录酶 - 聚合酶链反应（RT-PCR）分析表明，许多白血病CD34（+）G（0）细胞也表达了来自8个CML患者的BCR-ABL mRNA，CD34（+）也标记为羧基氟二甲酸二乙酸二乙酸酯二甲酸二甲酸酯二酰基二氨基二氨基二氨基二氨基二酰基二酰基二酰基二酰基二酰基二氨基二酰基二氨基二氨基二酰基二氨基二氨基二酰基二酰基二酰基二氨基二酰基。在培养之前（有或没有添加生长因子）之前，diester（cfse）允许保持静止领带的活细胞， CFSE+）在4天后被FAG检索后，在第二种策略分离出的所有静态种群中再次检测到白血病祖细胞，包括暴露于FLT3-ligand，钢质因子，Interleukin-3，Interleukin-3，Interleukin-6和颗粒细胞的组合的祖先。这些发现刺激菌落刺激因素，提供了第一个直接和明确的证据，证明患有体外和体内干细胞特性的CML患者的白血病细胞。 （c）1999年美国血液学学会。
Chronic myeloid leukemia (CML) is characterized by an increased proliferative activity of the leukemic progenitors that produce an elevated number of mature granulocytes. Nevertheless, cell cycle-active agents, even in very high doses, are alone unable to eradicate the leukemic clone, suggesting the presence of a rare subset of quiescent leukemic stem cells. To isolate such cells, we first used Hoechst 33342 and Pyronin Y staining to obtain viable G(0) and G(1)/S/G(2)/M fractions of CD34(+) cells by fluorescence-activated cell sorting (FACS) from 6 chronic-phase CML patients' samples and confirmed the quiescent and cycling status of the 2 fractions by demonstration of expected patterns of Ki-67 and D cyclin expression. Leukemic (Ph+/BCR-ABL(+)) cells with in vitro progenitor activity and capable of engrafting immunodeficient mice were identified in the directly isolated Go cells. Single-cell reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed that many leukemic CD34(+) G(0) cells also expressed BCR-ABL mRNA, CD34(+) from 8 CML patients were also labeled with carboxyfluorescein diacetate succinimidyl diester (CFSE) before being cultured (with and without added growth factors) to allow viable cells that had remained quiescent tie, CFSE+) after 4 days to be retrieved by FAGS, Leukemic progenitors were again detected in all quiescent populations isolated by this second strategy, including those exposed to a combination of flt3-ligand, Steel factor, interleukin-3, interleukin-6, and granulocyte colony-stimulating factor, These findings provide the first direct and definitive evidence of a deeply but reversibly quiescent subpopulation of leukemic cells in patients with CML with both in vitro and in vivo stem cell properties. (C) 1999 by The American Society of Hematology.