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HESC cryopreservation: use of dimethylsulfoxide and hydroxyethylstarch in an adapted protocol for efficient hESC banking

VERONIQUE T'JOEN UGent, Linde De Grande UGent, Heidi Declercq UGent and Maria Cornelissen UGent (2011) CYTOTHERAPY.
abstract
hESC are one of the most interesting cell types for tissue engineering purposes, advancements in drug screening, discoveries in basic scientific research and the conduction of developmental studies. One prerequisite for fast advancement in these areas is the availability of large amounts of well-characterized hESC cells from hESC banks. The banking of hESC for research and future clinical applications requires optimized cryopreservation and thawing protocols. We investigated several cryopreservation solutions and protocols for slow freezing. As our focus lies on future clinical applications of hESC cells and engineered constructs, we attempted to eliminate high amounts of cryoprotectors, serum and other added substances. Preliminary freezing tests were performed with the VUB01 cell line. Different concentrations of DMSO (dimethylsulfoxide), EG (ethylene glycol) and HES (hydroxyethylstarch) were examined in combination with two different application protocols. The use of 5% DMSO + 5% HES and the adapted protocol (treatment with Cell Dissociation Solution) led to a high increase in recovery rate, as can be seen in the figure 1. We also demonstrated a good and fast expansion of the cryopreserved hESC (quantitative and qualitative follow-up during 3 passages). Subsequently, the two best conditions (5% DMSO + 5% HES and 10% DMSO + 10% HES) were tested with 4 other hESC lines (181, H9, H1 and UGhent2) in order to establish the reproducibility of the cryopreservation conditions (figure 2). The adapted protocol with 5% DMSO and 5% HES remained the best condition for all cell lines and the recovery rates varied between 11,38% and 42,05%. We conclude that 5% DMSO + 5% HES in combination with the adapted protocol is the best way to cryopreserve hESC in hESC cell banks for future clinical applications.
Please use this url to cite or link to this publication:
author
organization
year
type
conference
publication status
in press
subject
keyword
cryopreservation, embryonic stem cells
in
CYTOTHERAPY
Cytotherapy
conference name
17th Annual ISCT meeting (ISCT - 2011)
conference location
Rotterdam, The Netherlands
conference start
2011-05-18
conference end
2011-05-21
JCR category
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
JCR impact factor
3.627 (2011)
JCR rank
33/157 (2011)
JCR quartile
1 (2011)
ISSN
1465-3249
language
English
UGent publication?
yes
classification
C3
id
1255586
handle
http://hdl.handle.net/1854/LU-1255586
date created
2011-06-07 09:47:08
date last changed
2011-06-23 15:32:25
@inproceedings{1255586,
  abstract     = {hESC are one of the most interesting cell types for tissue engineering purposes, advancements in drug screening, discoveries in basic scientific research and the conduction of developmental studies. One prerequisite for fast advancement in these areas is the availability of large amounts of well-characterized hESC cells from hESC banks. The banking of hESC for research and future clinical applications requires optimized cryopreservation and thawing protocols. We investigated several cryopreservation solutions and protocols for slow freezing. As our focus lies on future clinical applications of hESC cells and engineered constructs, we attempted to eliminate high amounts of cryoprotectors, serum and other added substances. Preliminary freezing tests were performed with the VUB01 cell line. Different concentrations of DMSO (dimethylsulfoxide), EG (ethylene glycol) and HES (hydroxyethylstarch) were examined in combination with two different application protocols. The use of 5\% DMSO + 5\% HES and the adapted protocol (treatment with Cell Dissociation Solution) led to a high increase in recovery rate, as can be seen in the figure 1. We also demonstrated a good and fast expansion of the cryopreserved hESC (quantitative and qualitative follow-up during 3 passages). Subsequently, the two best conditions (5\% DMSO + 5\% HES and 10\% DMSO + 10\% HES) were tested with 4 other hESC lines (181, H9, H1 and UGhent2) in order to establish the reproducibility of the cryopreservation conditions (figure 2). The adapted protocol with 5\% DMSO and 5\% HES remained the best condition for all cell lines and the recovery rates varied between 11,38\% and 42,05\%. We conclude that 5\% DMSO + 5\% HES in combination with the adapted protocol is the best way to cryopreserve hESC in hESC cell banks for future clinical applications.},
  author       = {T'JOEN, VERONIQUE and De Grande, Linde and Declercq, Heidi and Cornelissen, Maria},
  booktitle    = {CYTOTHERAPY},
  issn         = {1465-3249},
  keyword      = {cryopreservation,embryonic stem cells},
  language     = {eng},
  location     = {Rotterdam, The Netherlands},
  title        = {HESC cryopreservation: use of dimethylsulfoxide and hydroxyethylstarch in an adapted protocol for efficient hESC banking},
  year         = {2011},
}

Chicago
T’JOEN, VERONIQUE, Linde De Grande, Heidi Declercq, and Maria Cornelissen. 2011. “HESC Cryopreservation: Use of Dimethylsulfoxide and Hydroxyethylstarch in an Adapted Protocol for Efficient hESC Banking.” In Cytotherapy.
APA
T’JOEN, V., De Grande, L., Declercq, H., & Cornelissen, M. (2011). HESC cryopreservation: use of dimethylsulfoxide and hydroxyethylstarch in an adapted protocol for efficient hESC banking. CYTOTHERAPY. Presented at the 17th Annual ISCT meeting (ISCT - 2011).
Vancouver
1.
T’JOEN V, De Grande L, Declercq H, Cornelissen M. HESC cryopreservation: use of dimethylsulfoxide and hydroxyethylstarch in an adapted protocol for efficient hESC banking. CYTOTHERAPY. 2011.
MLA
T’JOEN, VERONIQUE, Linde De Grande, Heidi Declercq, et al. “HESC Cryopreservation: Use of Dimethylsulfoxide and Hydroxyethylstarch in an Adapted Protocol for Efficient hESC Banking.” Cytotherapy. 2011. Print.