Isolation of live label-retaining cells and cells undergoing asymmetric cell division via nonrandom chromosomal cosegregation from human cancers

Danielle Hari, Hong Wu Xin, Kshama Jaiswal, Gordon Wiegand, Bo Kyu Kim, Che Ambe, Douglas Burka, Tomotake Koizumi, Satyajit Ray, Susan Garfield, Snorri Thorgeirsson, Itzhak Avital

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)1649-1658
Number of pages10
JournalStem Cells and Development
Volume20
Issue number10
DOIs
StatePublished - 1 Oct 2011
Externally publishedYes

ASJC Scopus subject areas

  • Hematology
  • Developmental Biology
  • Cell Biology

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