title: Isolation of live label-retaining cells and cells undergoing asymmetric cell division via nonrandom chromosomal cosegregation from human cancers.
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.
authors.affiliation: Surgery Branch, Center for Cancer Research, National Cancer Institute, National Institute of Health, Bethesda, Maryland 20892, USA.
authors.forename: Danielle
authors.initials: D
authors.lastname: Hari
authors.forename: Hong-Wu
authors.initials: HW
authors.lastname: Xin
authors.forename: Kshama
authors.initials: K
authors.lastname: Jaiswal
authors.forename: Gordon
authors.initials: G
authors.lastname: Wiegand
authors.forename: Bo-Kyu
authors.initials: BK
authors.lastname: Kim
authors.forename: Che
authors.initials: C
authors.lastname: Ambe
authors.forename: Douglas
authors.initials: D
authors.lastname: Burka
authors.forename: Tomotake
authors.initials: T
authors.lastname: Koizumi
authors.forename: Satyajit
authors.initials: S
authors.lastname: Ray
authors.forename: Susan
authors.initials: S
authors.lastname: Garfield
authors.forename: Snorri
authors.initials: S
authors.lastname: Thorgeirsson
authors.forename: Itzhak
authors.initials: I
authors.lastname: Avital
mesh_terms: D060049:Asymmetric Cell Division; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D020090:Chromosome Segregation; D003116:Color; D006801:Humans; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology*; D009711:Nucleotides / Q000378:metabolism; D013194:Staining and Labeling
chemical_list: D009711:Nucleotides