DIFFERENTIATION OF HL-60 PROMYELOCYTIC LEUKEMIA-CELLS MONITORED BY FLOW CYTOMETRIC MEASUREMENT OF NITRO BLUE TETRAZOLIUM (NBT) REDUCTION
DIFFERENTIATION OF HL-60 PROMYELOCYTIC LEUKEMIA-CELLS MONITORED BY FLOW CYTOMETRIC MEASUREMENT OF NITRO BLUE TETRAZOLIUM (NBT) REDUCTION
复制标题
DOI：
10.1002/cyto.990060110
复制
发表时间：
1985-01-01
期刊：
影响因子：
--
通讯作者：
SARTORELLI, AC
中科院分区：
文献类型：
--
作者：
BLAIR, OC;CARBONE, R;SARTORELLI, AC
硝基蓝四唑 (NBT) 还原成不溶性蓝色甲臜颗粒是在成熟粒细胞刺激诱导的呼吸爆发过程中发生的，通常用作 HL-60 急性早幼粒细胞白血病细胞粒细胞分化程度的指标。通过流式细胞术(FCM)测量正向和90°来监测由二甲基亚砜(DMSO)或视黄酸诱导的HL-60白血病细胞的分化。 NBT处理细胞的光散射。 双参数相关分析允许区分前向增加和 90° 减小的细胞。光散射（NBT-），细胞前向减少，90°增加。光散射 (NBT+)。用 1% 多聚甲醛固定 NBT 处理的细胞有利于流式细胞术分析，并可对 NBT 减少的差异进行定量。与表现出分级反应的视黄酸处理的细胞相比，DMSO 诱导的细胞表达 NBT 全部或全部减少到甲臜。对碘化丙啶与 NBT 结合染色的 HL-60 白血病细胞进行三参数流式细胞术分析，可以确定 NBT 处理细胞的细胞周期分布。
Reduction of nitro blue tetrazolium (NBT) to insoluble blue formazan granules occurs during the stimulus-induced respiratory burst of mature granulocytes and is routinely used as an indicator of the extent of granulocytic differentiation of HL-60 acute promyelocytic leukemia cells. The differentiation of HL-60 leukemia cells induced by dimethylsulfoxide (DMSO) or retinoic acid was monitored by flow cytometric (FCM) measurement of forward and 90.degree. light scatter of NBT treated cells. Two-parameter correlated analysis is permitted a distinction between cells with increased forward and decreased 90.degree. light scatter (NBT-), and cells with decreased forward and increased 90.degree. light scatter (NBT+). Fixation of NBT treated cells with 1% paraformaldehyde facilitated flow cytometric analysis, and allowed differences in NBT reduction to be quantitated. DMSO-induced cells expressed an all-or-none reduction of NBT to formazan, compared with retinoic acid treated cells that exhibited a graded response. Three parameters flow cytometric analysis of HL-60 leukemia cells stained with propidium iodine in combination with NBT allowed the determination of the cell cycle distribution of NBT-treated cells.