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Serum replacement with a growth factor–free synthetic substance in culture medium contributes to effective establishment of mouse embryonic stem cells of various origins

Cited 8 time in Web of Science Cited 10 time in Scopus
Authors
Lee, Seung Tae; Oh, Se Woong; Kim, Dae-Yong; Han, Jae Yong; Moon, Shin Yong; Lim, Jeong Mook
Issue Date
2006-09-06
Publisher
Elsevier
Citation
Fertil Steril 2006;86:1137-45
Keywords
Mouseembryonic stem (ES) cellestablishmentserum replacementsingle-cell culture
Abstract
Objective: To evaluate whether serum replacement with growth factor–free synthetic substances contributed to
the effective establishment of embryonic stem (ES) cells.
Design: Randomized, prospective model study.
Setting: Gamete and stem cell biotechnology laboratory at Seoul National University in Korea.
Animal(s): F1 (C57BL6 DBA2) mice.
Intervention(s): Blastocysts of different origins were cultured in serum-replaced media.
Main Outcome Measure(s): Embryonic stem cell establishment.
Result(s): Eight batches of ES cells were established from colony-forming inner cell mass cells after the
replacement of fetal bovine serum (FBS) with synthetic knockout serum replacement (KSR) in mkDMEM. The
established cells were positive for ES cell markers and formed both embryoid bodies in vitro and teratomas in
vivo, but the established cell batches and control (transformed) ES cells responded differently to the culture
media. Higher levels of cell viability were detected after the replacement with the 75:25 FBS–KSR mixture than
with any other mixtures, and a gradual decrease in viability was detected as the KSR volume ratio was increased.
The 75:25 FBS–KSR mixture-containing medium supported ES cell establishment of outbred ICR, F1, and F2 of
C57BL6/DBA2; F1 parthenogenetic and ES cell–complemented tetraploid blastocysts; and single ES-cell
cultures.
Conclusion(s): A serum-replaced medium could be used for effective ES-cell establishment of various origins.
(Fertil Steril 2006;86(Suppl 3):1137– 45. ©2006 by American Society for Reproductive Medicine.)
Objective: To evaluate whether serum replacement with growth factor–free synthetic substances contributed to
the effective establishment of embryonic stem (ES) cells.
Design: Randomized, prospective model study.
Setting: Gamete and stem cell biotechnology laboratory at Seoul National University in Korea.
Animal(s): F1 (C57BL6 DBA2) mice.
Intervention(s): Blastocysts of different origins were cultured in serum-replaced media.
Main Outcome Measure(s): Embryonic stem cell establishment.
Result(s): Eight batches of ES cells were established from colony-forming inner cell mass cells after the
replacement of fetal bovine serum (FBS) with synthetic knockout serum replacement (KSR) in mkDMEM. The
established cells were positive for ES cell markers and formed both embryoid bodies in vitro and teratomas in
vivo, but the established cell batches and control (transformed) ES cells responded differently to the culture
media. Higher levels of cell viability were detected after the replacement with the 75:25 FBS–KSR mixture than
with any other mixtures, and a gradual decrease in viability was detected as the KSR volume ratio was increased.
The 75:25 FBS–KSR mixture-containing medium supported ES cell establishment of outbred ICR, F1, and F2 of
C57BL6/DBA2; F1 parthenogenetic and ES cell–complemented tetraploid blastocysts; and single ES-cell
cultures.
Conclusion(s): A serum-replaced medium could be used for effective ES-cell establishment of various origins.
(Fertil Steril 2006;86(Suppl 3):1137– 45. ©2006 by American Society for Reproductive Medicine.)
ISSN
0362-028X
Language
English
URI
https://hdl.handle.net/10371/6899
DOI
https://doi.org/10.1016/j.fertnstert.2006.01.056
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College of Veterinary Medicine (수의과대학)Dept. of Veterinary Medicine (수의학과)Journal Papers (저널논문_수의학과)
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