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Expansion of human embryonic stem cells: a comparative study

VERONIQUE T'JOEN UGent, Heidi Declercq UGent and Maria Cornelissen UGent (2011) CELL PROLIFERATION. 44(5). p.462-476
abstract
Objectives: Human embryonic stem cells (hESC) are promising for tissue engineering (TE) purposes due to their unique properties. However, current standard mechanical passaging techniques limit rates of possible TE experiments, as it is difficult to obtain high enough numbers of the cells for experimentation. In this study, several dissociative solutions and application methods are tested for their applicability to, and influence on, hESC culture and expansion. Materials and methods: Expansion of two hESC lines, H1 and VUB01, subjected to different passaging techniques, was evaluated. Four dissociative solutions - TrypLE (TM) Express, Trypsin-EDTA, Cell Dissociation Solution and Accutase (TM) - were combined with two application protocols. As reference conditions, manual and bead-based passaging techniques were used. Results: Results showed that use of Cell Dissociation Solution in combination with a slow adaptation protocol, generated the best expansion profile for both cell lines. The hESC single cell lines remained pluripotent, had good expansion profiles and were capable of differentiation into representatives of all three germ layers. Reproducibility of the results was confirmed by adaptation for three other hESC lines. Conclusion: Use of Cell Dissociation Solution, combined with slow adaptation protocol, allows a fast switch from the mechanical passaging technique to a single-cell split technique, generating stable and robust hESC cell lines, which allow for large scale expansion of hESC for TE purposes.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
MAINTENANCE, DERIVATION, CULTURE-CONDITIONS, FEEDER-FREE, LINES, CD30, EXPRESSION, EFFICIENT, DISSOCIATION, SURVIVAL
journal title
CELL PROLIFERATION
Cell Prolif.
volume
44
issue
5
pages
462 - 476
Web of Science type
Article
Web of Science id
000295128900008
JCR category
CELL BIOLOGY
JCR impact factor
2.521 (2011)
JCR rank
118/178 (2011)
JCR quartile
3 (2011)
ISSN
0960-7722
DOI
10.1111/j.1365-2184.2011.00773.x
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
1939208
handle
http://hdl.handle.net/1854/LU-1939208
date created
2011-11-07 11:30:38
date last changed
2011-11-07 14:18:15
@article{1939208,
  abstract     = {Objectives: Human embryonic stem cells (hESC) are promising for tissue engineering (TE) purposes due to their unique properties. However, current standard mechanical passaging techniques limit rates of possible TE experiments, as it is difficult to obtain high enough numbers of the cells for experimentation. In this study, several dissociative solutions and application methods are tested for their applicability to, and influence on, hESC culture and expansion.
Materials and methods: Expansion of two hESC lines, H1 and VUB01, subjected to different passaging techniques, was evaluated. Four dissociative solutions - TrypLE (TM) Express, Trypsin-EDTA, Cell Dissociation Solution and Accutase (TM) - were combined with two application protocols. As reference conditions, manual and bead-based passaging techniques were used.
Results: Results showed that use of Cell Dissociation Solution in combination with a slow adaptation protocol, generated the best expansion profile for both cell lines. The hESC single cell lines remained pluripotent, had good expansion profiles and were capable of differentiation into representatives of all three germ layers. Reproducibility of the results was confirmed by adaptation for three other hESC lines.
Conclusion: Use of Cell Dissociation Solution, combined with slow adaptation protocol, allows a fast switch from the mechanical passaging technique to a single-cell split technique, generating stable and robust hESC cell lines, which allow for large scale expansion of hESC for TE purposes.},
  author       = {T'JOEN, VERONIQUE and Declercq, Heidi and Cornelissen, Maria},
  issn         = {0960-7722},
  journal      = {CELL PROLIFERATION},
  keyword      = {MAINTENANCE,DERIVATION,CULTURE-CONDITIONS,FEEDER-FREE,LINES,CD30,EXPRESSION,EFFICIENT,DISSOCIATION,SURVIVAL},
  language     = {eng},
  number       = {5},
  pages        = {462--476},
  title        = {Expansion of human embryonic stem cells: a comparative study},
  url          = {http://dx.doi.org/10.1111/j.1365-2184.2011.00773.x},
  volume       = {44},
  year         = {2011},
}

Chicago
T’JOEN, VERONIQUE, Heidi Declercq, and Maria Cornelissen. 2011. “Expansion of Human Embryonic Stem Cells: a Comparative Study.” Cell Proliferation 44 (5): 462–476.
APA
T’JOEN, V., Declercq, H., & Cornelissen, M. (2011). Expansion of human embryonic stem cells: a comparative study. CELL PROLIFERATION, 44(5), 462–476.
Vancouver
1.
T’JOEN V, Declercq H, Cornelissen M. Expansion of human embryonic stem cells: a comparative study. CELL PROLIFERATION. 2011;44(5):462–76.
MLA
T’JOEN, VERONIQUE, Heidi Declercq, and Maria Cornelissen. “Expansion of Human Embryonic Stem Cells: a Comparative Study.” CELL PROLIFERATION 44.5 (2011): 462–476. Print.