| FEDR, Radek, Zuzana PERNICOVÁ, Eva SLABÁKOVÁ, Nicol STRAKOVÁ, Jan BOUCHAL, Michal GREPL, Alois KOZUBÍK a Karel SOUČEK. Automatic cell cloning assay for determining the clonogenic capacity of cancer and cancer stem-like cells. Cytometry Part A. John Wiley & Sons, Inc., 2013, roč. 83A, č. 5, s. 472-482. ISSN 1552-4922. Dostupné z: https://dx.doi.org/10.1002/cyto.a.22273. |
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@article{1095411,
author = {Fedr, Radek and Pernicová, Zuzana and Slabáková, Eva and Straková, Nicol and Bouchal, Jan and Grepl, Michal and Kozubík, Alois and Souček, Karel},
article_number = {5},
doi = {http://dx.doi.org/10.1002/cyto.a.22273},
keywords = {clonogenic assay; clonogenic capacity; plating efficiency; cancer stem cells; extreme limiting dilution analysis; single cell sorting},
language = {eng},
issn = {1552-4922},
journal = {Cytometry Part A},
title = {Automatic cell cloning assay for determining the clonogenic capacity of cancer and cancer stem-like cells},
volume = {83A},
year = {2013}
}
TY - JOUR
ID - 1095411
AU - Fedr, Radek - Pernicová, Zuzana - Slabáková, Eva - Straková, Nicol - Bouchal, Jan - Grepl, Michal - Kozubík, Alois - Souček, Karel
PY - 2013
TI - Automatic cell cloning assay for determining the clonogenic capacity of cancer and cancer stem-like cells
JF - Cytometry Part A
VL - 83A
IS - 5
SP - 472-482
EP - 472-482
PB - John Wiley & Sons, Inc.
SN - 15524922
KW - clonogenic assay
KW - clonogenic capacity
KW - plating efficiency
KW - cancer stem cells
KW - extreme limiting dilution analysis
KW - single cell sorting
N2 - The clonogenic assay is a well-established in vitro method for testing the survival and proliferative capability of cells. It can be used to determine the cytotoxic effects of various treatments including chemotherapeutics and ionizing radiation. However, this approach can also characterize cells with different phenotypes and biological properties, such as stem cells or cancer stem cells. In this study, we implemented a faster and more precise method for assessing the cloning efficiency of cancer stem-like cells that were characterized and separated using a high-speed cell sorter. Cell plating onto a microplate using an automatic cell deposition unit was performed in a single-cell or dilution rank mode by the fluorescence-activated cell sorting method. We tested the new automatic cell-cloning assay (ACCA) on selected cancer cell lines and compared it with the manual approach. The obtained results were also compared with the results of the limiting dilution assay for different cell lines. We applied the ACCA to analyze the cloning capacity of different subpopulations of prostate and colon cancer cells based on the expression of the characteristic markers of stem (CD44 and CD133) and cancer stem cells (TROP-2, CD49f, and CD44). Our results revealed that the novel ACCA is a straightforward approach for determining the clonogenic capacity of cancer stem-like cells identified in both cell lines and patient samples.
ER -
FEDR, Radek, Zuzana PERNICOVÁ, Eva SLABÁKOVÁ, Nicol STRAKOVÁ, Jan BOUCHAL, Michal GREPL, Alois KOZUBÍK a Karel SOUČEK. Automatic cell cloning assay for determining the clonogenic capacity of cancer and cancer stem-like cells. \textit{Cytometry Part A}. John Wiley \&{} Sons, Inc., 2013, roč.~83A, č.~5, s.~472-482. ISSN~1552-4922. Dostupné z: https://dx.doi.org/10.1002/cyto.a.22273.
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