RAPID DECLINE OF CHRONIC MYELOID LEUKEMIC-CELLS IN LONG-TERM CULTURE DUE TO A DEFECT AT THE LEUKEMIC STEM-CELL LEVEL
RAPID DECLINE OF CHRONIC MYELOID LEUKEMIC-CELLS IN LONG-TERM CULTURE DUE TO A DEFECT AT THE LEUKEMIC STEM-CELL LEVEL
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DOI：
10.1073/pnas.89.13.6192
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发表时间：
1992-07-01
影响因子：
11.1
通讯作者：
EAVES, AC
中科院分区：
文献类型：
--
作者：
UDOMSAKDI, C;EAVES, CJ;EAVES, AC
在这份报告中，我们描述了一个定量的体外试验，最原始的类型白血病的前体尚未定义在慢性粒细胞白血病（CML）患者。该试验基于最近描述的原始正常人造血细胞的“长期培养起始细胞”（LTC-IC）试验。当与感受态成纤维细胞饲养层共培养时，这些细胞在至少5周后产生在次级半固体测定培养物中可检测到的多个单系和多系克隆生成祖细胞。通过将来自患者的高度富集的白血病细胞源接种到正常饲养细胞上而启动的类似培养物显示，5周后的集落形成细胞输出与输入接种物在低至有限数量的输入细胞的宽范围内线性相关，从而允许使用标准有限稀释分析技术来确定白血病LTC-IC的绝对频率。发现CML骨髓中的白血病LTC-IC浓度降低，在CML血液中平均降低至10（5）倍。通过与正常血液或骨髓LTC-IC值比较，评估每个白血病LTC-IC产生的克隆原细胞数，显示该功能在白血病LTC-IC中未改变[即，3.1每个CML LTC-IC +/- 0.4个克隆原细胞（平均值+/- SEM，n = 6），而正常血液和骨髓LTC-IC分别为3.7 +/- 1.2（n = 3）和4.3 +/- 0.4（n = 5）]。与此相反，白血病LTC-IC维持在LTC被证明是高度缺陷相比，正常的LTC-IC的血液或骨髓来源。因此，当将原代LTC的细胞继代培养至二次LTC-IC试验中时，白血病LTC-IC在培养的前10天内迅速下降（>30倍），而正常LTC-IC数量在此期间保持不变。这些发现说明了原始人类造血细胞中的自我维持和分化事件如何通过致癌过程进行差异调节，并为进一步研究其操纵，分析和治疗开发提供了框架。
In this report we describe a quantitative in vitro assay for the most primitive type of leukemic precursors yet defined in patients with chronic myeloid leukemia (CML). This assay is based on the recently described "long-term culture-initiating cell" (LTC-IC) assay for primitive normal human hematopoietic cells. Such cells, when cocultured with competent fibroblast feeder layers, give rise after a minimum of 5 weeks to multiple single and multilineage clonogenic progenitors detectable in secondary semisolid assay cultures. Similar cultures initiated by seeding a highly enriched source of leukemic cells from patients onto normal feeders showed the clonogenic cell output after 5 weeks to be linearly related to the input innoculum over a wide range down to limiting numbers of input cells, thus allowing absolute frequencies of leukemic LTC-ICs to be determined using standard limiting dilution analysis techniques. Leukemic LTC-IC concentrations in CML marrow were found to be decreased, on average to 10(5) times) in CML blood. Assessment of the number of clonogenic cells produced per leukemic LTC-IC by comparison to normal blood or marrow LTC-IC values showed this function to be unchanged in leukemic LTC-ICs [i.e., 3.1 +/- 0.4 clonogenic cells per CML LTC-IC (mean +/- SEM, n = 6) versus 3.7 +/- 1.2 (n = 3) and 4.3 +/- 0.4 (n = 5), respectively, for normal blood and marrow LTC-ICs]. In contrast, leukemic LTC-IC maintenance in LTC proved to be highly defective by comparison to normal LTC-IC of either blood or marrow origin. Thus, when cells from primary LTC were subcultured into secondary LTC-IC assays, leukemic LTC-IC rapidly declined (>30-fold) within the first 10 days of culture, whereas normal LTC-IC numbers remained unchanged during this period. These findings illustrate how self-maintenance and differentiation events in primitive human hematopoietic cells can be differentially modulated by an oncogenic process and provide a framework for further studies of their manipulation, analysis, and therapeutic exploitation.