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Efficient large scale expansion of human embryonic stem cells: comparison of 4 different solutions for single cell passaging

VERONIQUE T'JOEN UGent, Heidi Declercq UGent and Maria Cornelissen UGent (2010) 3rd annual stem cells Europe conference, Abstracts.
abstract
Human embryonic stem cells (hESC) are promising for tissue engineering (TE) purposes due to their unique properties. However, the current standard mechanical passaging technique limits the rate of possible TE experiments. Alternative single cell passaging techniques have been described in literature with varying results. In this comparative study, the H1 and VUB02 hESC lines were tested in combination with a fast and slow adaptation protocol and 4 different dissociative solutions to determine the best strategy to quickly obtain large amounts of stable hESC. The following dissociative solutions were examined: TrypLE™ Express, Trypsin-EDTA, Cell Dissociation Solution and Accutase™. Results showed that it was possible to generate pluripotent hESC colonies using all 4 dissociative solutions. The use of Cell Dissociation Solution proved to be the most efficient, as it generated the most colonies and sustained a high and stable expansion rate for both hESC cell lines. Application of the slow adaptation protocol resulted in an overall higher and quicker expansion rate than when the fast adaptation protocol was applied. Additionally, Cell Dissociation Solution is an enzyme- and animal-free solution, which makes these results more interesting regarding future clinical applications. The obtained hESC cell lines maintained their pluripotent capacity and had a stable karyotypic profile. We conclude that Cell dissociation solution in combination with a slow adaptation protocol allows a fast switch from the traditional mechanical passaging technique to a single cells split technique, generating a stable and robust hESC cell line, which allows large scale expansion for TE purposes.
Please use this url to cite or link to this publication:
author
organization
year
type
conference
publication status
unpublished
subject
keyword
expansion, embryonic stem cells, single cell culture
in
3rd annual stem cells Europe conference, Abstracts
conference name
3rd Annual Stem cells Europe Conference
conference location
Edinburgh, Scotland
conference start
2010-08-24
conference end
2010-08-25
language
English
UGent publication?
yes
classification
C3
id
1254197
handle
http://hdl.handle.net/1854/LU-1254197
alternative location
http://mms.technologynetworks.net/posters/0791.pdf
date created
2011-06-06 15:56:40
date last changed
2011-06-23 15:27:05
@inproceedings{1254197,
  abstract     = {Human embryonic stem cells (hESC) are promising for tissue engineering (TE) purposes due to their unique properties. However, the current standard mechanical passaging technique limits the rate of possible TE experiments. Alternative single cell passaging techniques have been described in literature with varying results. In this comparative study, the H1 and VUB02 hESC lines were tested in combination with a fast and slow adaptation protocol and 4 different dissociative solutions to determine the best strategy to quickly obtain large amounts of stable hESC. The following dissociative solutions were examined: TrypLE{\texttrademark} Express, Trypsin-EDTA, Cell Dissociation Solution and Accutase{\texttrademark}. Results showed that it was possible to generate pluripotent hESC colonies using all 4 dissociative solutions. The use of Cell Dissociation Solution proved to be the most efficient, as it generated the most colonies and sustained a high and stable expansion rate for both hESC cell lines. Application of the slow adaptation protocol resulted in an overall higher and quicker expansion rate than when the fast adaptation protocol was applied. Additionally, Cell Dissociation Solution is an enzyme- and animal-free solution, which makes these results more interesting regarding future clinical applications. The obtained hESC cell lines maintained their pluripotent capacity and had a stable karyotypic profile. We conclude that Cell dissociation solution in combination with a slow adaptation protocol allows a fast switch from the traditional mechanical passaging technique to a single cells split technique, generating a stable and robust hESC cell line, which allows large scale expansion for TE purposes.},
  author       = {T'JOEN, VERONIQUE and Declercq, Heidi and Cornelissen, Maria},
  booktitle    = {3rd annual stem cells Europe conference, Abstracts},
  keyword      = {expansion,embryonic stem cells,single cell culture},
  language     = {eng},
  location     = {Edinburgh, Scotland},
  title        = {Efficient large scale expansion of human embryonic stem cells: comparison of 4 different solutions for single cell passaging},
  url          = {http://mms.technologynetworks.net/posters/0791.pdf},
  year         = {2010},
}

Chicago
T’JOEN, VERONIQUE, Heidi Declercq, and Maria Cornelissen. 2010. “Efficient Large Scale Expansion of Human Embryonic Stem Cells: Comparison of 4 Different Solutions for Single Cell Passaging.” In 3rd Annual Stem Cells Europe Conference, Abstracts.
APA
T’JOEN, V., Declercq, H., & Cornelissen, M. (2010). Efficient large scale expansion of human embryonic stem cells: comparison of 4 different solutions for single cell passaging. 3rd annual stem cells Europe conference, Abstracts. Presented at the 3rd Annual Stem cells Europe Conference.
Vancouver
1.
T’JOEN V, Declercq H, Cornelissen M. Efficient large scale expansion of human embryonic stem cells: comparison of 4 different solutions for single cell passaging. 3rd annual stem cells Europe conference, Abstracts. 2010.
MLA
T’JOEN, VERONIQUE, Heidi Declercq, and Maria Cornelissen. “Efficient Large Scale Expansion of Human Embryonic Stem Cells: Comparison of 4 Different Solutions for Single Cell Passaging.” 3rd Annual Stem Cells Europe Conference, Abstracts. 2010. Print.