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An efficient, economical slow-freezing method for large-scale human embryonic stem cell banking

VERONIQUE T'JOEN UGent, Linde De Grande UGent, Heidi Declercq UGent and Maria Cornelissen UGent (2012) STEM CELLS AND DEVELOPMENT. 21(5). p.721-728
abstract
Human embryonic stem cells (hESCs) are one of the most interesting cell types for tissue engineering, cell therapy, basic scientific research, and drug screening. Fast advancement in these areas requires the availability of large amounts of safe and well-characterized hESCs from hESC banks. Therefore, optimized freezing protocols, allowing the cryopreservation of large amounts of hESC without direct contact with liquid nitrogen, need to be established. In this study, 6 different cryoprotector combinations [dimethylsulfoxide (DMSO), ethylene glycol, and hydroxyethylstarch (HES)] combined with 2 different application methods were screened with the VUB01 cell line, to establish a new slow-freezing protocol with high recovery rates and a good expansion capacity. Our best conditions were confirmed in 4 other hESC lines: H1, H9, 181, and UGent2. To our knowledge, this is the first time that HES is evaluated as a cryoprotector for hESCs. The use of 5% DMSO + 5% HES combined with a new detachment protocol leads to efficient hESC cryopreservation. This protocol involves treating the hESC colonies with cell dissociation solution, a mild dissociation solution uncommonly used for hESC culture. A recovery ratio ranging from 45.5% to 168.2% was obtained, and these were significantly different from the other tested conditions (Student's t-test, P < 0.05). The cryopreserved hESCs were morphologically comparable to control cells, exhibited a good expansion profile, were positive for pluripotent expression markers, and could still differentiate into the 3 germ layers. This new protocol allows efficient and economical hESC cryopreservation, ideal for hESC banking.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
LINES, SURVIVAL, BULK VITRIFICATION, ROCK INHIBITOR, CRYOPRESERVATION METHOD, STORAGE
journal title
STEM CELLS AND DEVELOPMENT
Stem Cells Dev.
volume
21
issue
5
pages
721 - 728
Web of Science type
Article
Web of Science id
000301292100008
JCR category
TRANSPLANTATION
JCR impact factor
4.67 (2012)
JCR rank
3/26 (2012)
JCR quartile
1 (2012)
ISSN
1547-3287
DOI
10.1089/scd.2011.0192
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
2940868
handle
http://hdl.handle.net/1854/LU-2940868
date created
2012-06-28 10:16:44
date last changed
2012-07-10 12:06:40
@article{2940868,
  abstract     = {Human embryonic stem cells (hESCs) are one of the most interesting cell types for tissue engineering, cell therapy, basic scientific research, and drug screening. Fast advancement in these areas requires the availability of large amounts of safe and well-characterized hESCs from hESC banks. Therefore, optimized freezing protocols, allowing the cryopreservation of large amounts of hESC without direct contact with liquid nitrogen, need to be established. In this study, 6 different cryoprotector combinations [dimethylsulfoxide (DMSO), ethylene glycol, and hydroxyethylstarch (HES)] combined with 2 different application methods were screened with the VUB01 cell line, to establish a new slow-freezing protocol with high recovery rates and a good expansion capacity. Our best conditions were confirmed in 4 other hESC lines: H1, H9, 181, and UGent2. To our knowledge, this is the first time that HES is evaluated as a cryoprotector for hESCs. The use of 5\% DMSO + 5\% HES combined with a new detachment protocol leads to efficient hESC cryopreservation. This protocol involves treating the hESC colonies with cell dissociation solution, a mild dissociation solution uncommonly used for hESC culture. A recovery ratio ranging from 45.5\% to 168.2\% was obtained, and these were significantly different from the other tested conditions (Student's t-test, P {\textlangle} 0.05). The cryopreserved hESCs were morphologically comparable to control cells, exhibited a good expansion profile, were positive for pluripotent expression markers, and could still differentiate into the 3 germ layers. This new protocol allows efficient and economical hESC cryopreservation, ideal for hESC banking.},
  author       = {T'JOEN, VERONIQUE and De Grande, Linde and Declercq, Heidi and Cornelissen, Maria},
  issn         = {1547-3287},
  journal      = {STEM CELLS AND DEVELOPMENT},
  keyword      = {LINES,SURVIVAL,BULK VITRIFICATION,ROCK INHIBITOR,CRYOPRESERVATION METHOD,STORAGE},
  language     = {eng},
  number       = {5},
  pages        = {721--728},
  title        = {An efficient, economical slow-freezing method for large-scale human embryonic stem cell banking},
  url          = {http://dx.doi.org/10.1089/scd.2011.0192},
  volume       = {21},
  year         = {2012},
}

Chicago
T’JOEN, VERONIQUE, Linde De Grande, Heidi Declercq, and Maria Cornelissen. 2012. “An Efficient, Economical Slow-freezing Method for Large-scale Human Embryonic Stem Cell Banking.” Stem Cells and Development 21 (5): 721–728.
APA
T’JOEN, V., De Grande, L., Declercq, H., & Cornelissen, M. (2012). An efficient, economical slow-freezing method for large-scale human embryonic stem cell banking. STEM CELLS AND DEVELOPMENT, 21(5), 721–728.
Vancouver
1.
T’JOEN V, De Grande L, Declercq H, Cornelissen M. An efficient, economical slow-freezing method for large-scale human embryonic stem cell banking. STEM CELLS AND DEVELOPMENT. 2012;21(5):721–8.
MLA
T’JOEN, VERONIQUE, Linde De Grande, Heidi Declercq, et al. “An Efficient, Economical Slow-freezing Method for Large-scale Human Embryonic Stem Cell Banking.” STEM CELLS AND DEVELOPMENT 21.5 (2012): 721–728. Print.