Isolation of Live Label-Retaining Cells and Cells Undergoing Asymmetric Cell Division via Nonrandom Chromosomal Cosegregation from Human Cancers
Isolation of Live Label-Retaining Cells and Cells Undergoing Asymmetric Cell Division via Nonrandom Chromosomal Cosegregation from Human Cancers
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DOI：
10.1089/scd.2010.0455
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发表时间：
2011-10-01
影响因子：
4
通讯作者：
Avital, Itzhak
中科院分区：
文献类型：
--
作者：
Hari, Danielle;Xin, Hong-Wu;Avital, Itzhak
随着时间的推移保留 DNA 标签的能力被认为与成体干细胞有关。最近，在癌症中发现了标签保留细胞（LRC）。 LRC 被认为是慢循环或非随机染色体共分离（ACD-NRCC）的不对称细胞分裂的结果。 ACD-NRCC 旨在将较旧的模板 DNA 链分离到子代干细胞中，并将新合成的 DNA 分离到用于分化的子细胞中。经历 ACD-NRCC 的细胞是否存在以及 LRC 的干细胞性质仍然存在争议。目前，为了检测LRC和ACD-NRCC，需要对细胞进行固定。因此，测试 LRC 和经历 ACD-NRCC 的细胞的干细胞性质和其他功能特征受到限制。在这里，我们展示了一种在活人肝癌细胞中用单色和双色核苷酸标记 DNA 的方法，无需固定。我们描述了一种通过荧光激活细胞分选和共聚焦显微镜验证来分离活 LRC 和经历 ACD-NRCC 的细胞的新方法。这有可能成为干细胞和癌症研究的强大辅助手段。
The ability to retain DNA labels over time is a property proposed to be associated with adult stem cells. Recently, label retaining cells (LRC) were indentified in cancer. LRC were suggested to be the result of either slow-cycling or asymmetric-cell-division with nonrandom-chromosomal-cosegregation (ACD-NRCC). ACD-NRCC is proposed to segregate the older template DNA strands into daughter stem cells and newly synthesized DNA into daughter cells destined for differentiation. The existence of cells undergoing ACD-NRCC and the stem-like nature of LRC remain controversial. Currently, to detect LRC and ACD-NRCC, cells need to undergo fixation. Therefore, testing the stem-cell nature and other functional traits of LRC and cells undergoing ACD-NRCC has been limited. Here, we show a method for labeling DNA with single and dual-color nucleotides in live human liver cancer cells avoiding the need for fixation. We describe a novel methodology for both the isolation of live LRC and cells undergoing ACD-NRCC via fluorescence-activated cell sorting with confocal microscopy validation. This has the potential to be a powerful adjunct to stem-cell and cancer research.