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The XTT cell proliferation assay applied to cell layers embedded in three-dimensional matrix

LYNN HUYCK UGent, Christophe Ampe UGent and Marleen Van Troys UGent (2012) ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES. 10(4). p.382-392
abstract
Abstract Cell proliferation, a main target in cancer therapy, is influenced by the surrounding three-dimensional (3D) extracellular matrix (ECM). In vitro drug screening is, thus, optimally performed under conditions in which cells are grown (embedded or trapped) in dense 3D matrices, as these most closely mimic the adhesive and mechanical properties of natural ECM. Measuring cell proliferation under these conditions is, however, technically more challenging compared with two-dimensional (2D) culture and other "3D culture conditions," such as growth on top of a matrix (pseudo-3D) or in spongy scaffolds with large pore sizes. Consequently, such measurements are only slowly applied on a wider scale. To advance this, we report on the equal quality (dynamic range, background, linearity) of measuring the proliferation of cell layers embedded in dense 3D matrices (collagen, Matrigel) compared with cells in 2D culture using the easy (one-step) and in 2D well-validated, 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT)-assay. The comparison stresses the differences in proliferation kinetics and drug sensitivity of matrix-embedded cells versus 2D culture. Using the specific cell-layer-embedded 3D matrix setup, quantitative measurements of cell proliferation and cell invasion are shown to be possible in similar assay conditions, and cytostatic, cytotoxic, and anti-invasive drug effects can thus be reliably determined and compared in physiologically relevant settings. This approach in the 3D matrix holds promise for improving early-stage, high-throughput drug screening, targeting either highly invasive or highly proliferative subpopulations of cancers or both.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
IN-VITRO, MESENCHYMAL STEM-CELLS, ENDOTHELIAL-CELLS, DRUG DISCOVERY, CULTURE MODELS, CYTOTOXICITY, GROWTH, MIGRATION, SCAFFOLD, MECHANISM
journal title
ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES
Assay Drug Dev. Technol.
volume
10
issue
4
pages
382 - 392
Web of Science type
Article
Web of Science id
000307864400008
JCR category
PHARMACOLOGY & PHARMACY
JCR impact factor
1.896 (2012)
JCR rank
160/259 (2012)
JCR quartile
3 (2012)
ISSN
1540-658X
DOI
10.1089/adt.2011.391
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
2130533
handle
http://hdl.handle.net/1854/LU-2130533
date created
2012-06-04 11:25:26
date last changed
2012-09-20 11:09:03
@article{2130533,
  abstract     = {Abstract Cell proliferation, a main target in cancer therapy, is influenced by the surrounding three-dimensional (3D) extracellular matrix (ECM). In vitro drug screening is, thus, optimally performed under conditions in which cells are grown (embedded or trapped) in dense 3D matrices, as these most closely mimic the adhesive and mechanical properties of natural ECM. Measuring cell proliferation under these conditions is, however, technically more challenging compared with two-dimensional (2D) culture and other {\textacutedbl}3D culture conditions,{\textacutedbl} such as growth on top of a matrix (pseudo-3D) or in spongy scaffolds with large pore sizes. Consequently, such measurements are only slowly applied on a wider scale. To advance this, we report on the equal quality (dynamic range, background, linearity) of measuring the proliferation of cell layers embedded in dense 3D matrices (collagen, Matrigel) compared with cells in 2D culture using the easy (one-step) and in 2D well-validated, 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT)-assay. The comparison stresses the differences in proliferation kinetics and drug sensitivity of matrix-embedded cells versus 2D culture. Using the specific cell-layer-embedded 3D matrix setup, quantitative measurements of cell proliferation and cell invasion are shown to be possible in similar assay conditions, and cytostatic, cytotoxic, and anti-invasive drug effects can thus be reliably determined and compared in physiologically relevant settings. This approach in the 3D matrix holds promise for improving early-stage, high-throughput drug screening, targeting either highly invasive or highly proliferative subpopulations of cancers or both.},
  author       = {HUYCK, LYNN and Ampe, Christophe and Van Troys, Marleen},
  issn         = {1540-658X},
  journal      = {ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES},
  keyword      = {IN-VITRO,MESENCHYMAL STEM-CELLS,ENDOTHELIAL-CELLS,DRUG DISCOVERY,CULTURE MODELS,CYTOTOXICITY,GROWTH,MIGRATION,SCAFFOLD,MECHANISM},
  language     = {eng},
  number       = {4},
  pages        = {382--392},
  title        = {The XTT cell proliferation assay applied to cell layers embedded in three-dimensional matrix},
  url          = {http://dx.doi.org/10.1089/adt.2011.391},
  volume       = {10},
  year         = {2012},
}

Chicago
Huyck, Lynn, Christophe Ampe, and Marleen Van Troys. 2012. “The XTT Cell Proliferation Assay Applied to Cell Layers Embedded in Three-dimensional Matrix.” Assay and Drug Development Technologies 10 (4): 382–392.
APA
Huyck, L., Ampe, C., & Van Troys, M. (2012). The XTT cell proliferation assay applied to cell layers embedded in three-dimensional matrix. ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES, 10(4), 382–392.
Vancouver
1.
Huyck L, Ampe C, Van Troys M. The XTT cell proliferation assay applied to cell layers embedded in three-dimensional matrix. ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES. 2012;10(4):382–92.
MLA
Huyck, Lynn, Christophe Ampe, and Marleen Van Troys. “The XTT Cell Proliferation Assay Applied to Cell Layers Embedded in Three-dimensional Matrix.” ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES 10.4 (2012): 382–392. Print.