Purification of leukemic blast cells from blood smears using laser microdissection.
Purification of leukemic blast cells from blood smears using laser microdissection.
复制标题
使用激光显微切割从血涂片中纯化白血病母细胞。
DOI：
10.1007/s12185-017-2227-z
复制
发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Adachi S.
中科院分区：
文献类型：
--
作者：
Matsuo H;Shiga S;Imai T;Kamikubo Y;Toki T;Terui K;Ito E;Adachi S.
在急性髓细胞白血病（AML）的治疗中，预后因素，包括基因突变和异常基因表达，使患者的风险分层。然而，在白血病母细胞比例很小的情况下，例如与唐氏综合征相关的AML（AML-DS），由于正常细胞比例很大，因此不可能精确检查预后因素。在这里，我们提出了一种新的方法来检查预后因素，从少量的诊断标本，并收集高纯度的白血病原始细胞的激光显微切割（LMD）的膜载玻片上的涂片。我们使用10%KPAM1细胞系悬浮液和含有20%原始细胞的外周血验证了该方法的有效性，所述原始细胞来自患有暂时性异常骨髓生成（TAM）的患者。在制备血液涂片后，收集约100个细胞并通过直接测序进行分析。在每个样本中检测到GATA-1的移码突变（分别为2 bp缺失和17 bp重复），表明KPAM 1和TAM原始细胞被准确纯化。这种新方法使我们能够在许多情况下精确地检查预后因素，即使是在白血病母细胞比例很小或需要保存的样本很小的情况下。
In treatment of acute myeloid leukemia (AML), prognostic factors, including gene mutation and abnormal gene expression, enable risk stratification of patients. However, in the case of a small proportion of leukemic blast cells, such as AML associated with Down syndrome (AML-DS), it is not possible to examine prognostic factors precisely due to the large proportion of normal cells. Here, we present a novel method for examining prognostic factors by making a smear on a membrane slide glass from a small amount of diagnostic specimen and collecting highly pure leukemic blast cells by laser microdissection (LMD). We verified the effectiveness of this method using 10% KPAM1 cell line suspension and peripheral blood containing 20% blast cells obtained from a patient with transient abnormal myelopoiesis (TAM). After making blood smears, approximately 100 cells were collected and analyzed by direct sequencing. Frameshift mutations (2 bp deletion and 17 bp duplication, respectively) inGATA-1were detected in each sample, suggesting KPAM1 and TAM blast cells were accurately purified. This novel method enables us to precisely examine prognostic factors in many cases, even in cases with a small proportion of leukemic blast cells or small specimens to preserve.