Walter H, Krob E J, Al-Romaihi F A, Johnson D, Lozzio C B
Laboratory of Chemical Biology, Veterans Administration Medical Center, Long Beach 90822.
Cell Biophys. 1988 Dec;13(3):173-87. doi: 10.1007/BF02918374.
The K-562 cell line is a culture of human leukemia stem cells originally derived from a patient with chronic myelogenous leukemia in blast crisis. We have applied a sensitive method capable of detecting subtle differences in charge-associated and noncharge-related cell surface properties between closely related cell populations to K-562 cells from different sources and having different histories. The method consists of isotopically labeling aliquots of each of two cell populations to be compared with 51Cr-chromate and mixing the labeled cells with an excess of unlabeled cells with which they are to be compared. The mixtures are subjected to countercurrent distribution in either a charge-sensitive or a noncharge-sensitive dextran-poly(ethylene glycol) aqueous two-phase system. The distribution curves are analyzed for total cells (in terms of electronic counts) and labeled cells (in terms of cpm). Alterations in relative specific activities through the distribution curves are indicative of differences in surface properties between such cell populations. Using this method we have found surface differences, both charge-associated and noncharge-related, between any two K-562 cell sublines examined. Interestingly, whereas we observed differences among K-562 sublines, we never witnessed a change in surface properties of the respective sublines. The differences among the sublines examined remained unaltered for more than 40 passages in our hands. It thus appears likely that the event(s) leading to an altered K-562 cell surface, detectable by partitioning, does not occur gradually.
K-562细胞系是一种人类白血病干细胞培养物,最初源自一名处于急变期的慢性粒细胞白血病患者。我们已将一种能够检测密切相关细胞群体之间电荷相关和非电荷相关细胞表面特性细微差异的灵敏方法应用于来自不同来源且具有不同历史的K-562细胞。该方法包括用51Cr-铬酸盐对要比较的两个细胞群体的等分试样进行同位素标记,并将标记的细胞与过量的未标记细胞混合,这些未标记细胞是它们要与之比较的对象。混合物在电荷敏感或非电荷敏感的葡聚糖-聚(乙二醇)水两相系统中进行逆流分配。分析分配曲线中的总细胞数(以电子计数表示)和标记细胞数(以每分钟计数表示)。通过分配曲线的相对比活性变化表明这些细胞群体之间表面特性的差异。使用这种方法,我们在任何两个检测的K-562细胞亚系之间发现了电荷相关和非电荷相关的表面差异。有趣的是,虽然我们在K-562亚系之间观察到差异,但我们从未目睹过各个亚系表面特性的变化。在我们手中,所检测的亚系之间的差异在超过40代中保持不变。因此,似乎导致可通过分配检测到的K-562细胞表面改变的事件不是逐渐发生的。