TY - JOUR
T1 - Isolation of live label-retaining cells and cells undergoing asymmetric cell division via nonrandom chromosomal cosegregation from human cancers
AU - Hari, Danielle
AU - Xin, Hong Wu
AU - Jaiswal, Kshama
AU - Wiegand, Gordon
AU - Kim, Bo Kyu
AU - Ambe, Che
AU - Burka, Douglas
AU - Koizumi, Tomotake
AU - Ray, Satyajit
AU - Garfield, Susan
AU - Thorgeirsson, Snorri
AU - Avital, Itzhak
PY - 2011/10/1
Y1 - 2011/10/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=80053410661&partnerID=8YFLogxK
U2 - 10.1089/scd.2010.0455
DO - 10.1089/scd.2010.0455
M3 - Article
C2 - 21294632
AN - SCOPUS:80053410661
SN - 1547-3287
VL - 20
SP - 1649
EP - 1658
JO - Stem Cells and Development
JF - Stem Cells and Development
IS - 10
ER -