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

Lee, Seung Tae; Oh, Se Woong; Kim, Dae Yong; Han, Jae Yong; Moon, Shin Yong; Lim, Jeong Mook

doi:10.1016/j.fertnstert.2006.01.056

« Previous Next »Fertility and Sterility
Volume 86, Issue 4, Supplement , Pages 1137-1145, October 2006

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

Seung Tae Lee, Ph.D.
- Department of Food and Animal Biotechnology, Seoul National University, Seoul, Korea
Se Woong Oh, D.V.M., M.S.
- College of Veterinary Medicine, Seoul National University, Seoul, Korea
Dae Yong Kim, D.V.M., Ph.D.
- College of Veterinary Medicine, Seoul National University, Seoul, Korea
Jae Yong Han, Ph.D.
- Department of Food and Animal Biotechnology, Seoul National University, Seoul, Korea
Shin Yong Moon, M.D., Ph.D.
- Stem Cell Research Center, Seoul National University, Seoul, Korea
- Medical Research Center, College of Medicine, Seoul National University, Seoul, Korea
Jeong Mook Lim, D.V.M., Ph.D.
- Department of Food and Animal Biotechnology, Seoul National University, Seoul, Korea
- Stem Cell Research Center, Seoul National University, Seoul, Korea
- Reprint requests: Jeong Mook Lim, D.V.M., Ph.D., Department of Food and Animal Biotechnology, Laboratory of Gamete and Stem Cell Biotechnology, Seoul National University, Building 200-#4223, Sillim-9 Dong, Seoul 151-921, Korea (FAX: 822-874-2555).

Received 5 October 2005; received in revised form 6 January 2006; accepted 6 January 2006. published online 04 September 2006.

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.

Key Words: Mouse , embryonic stem (ES) cell , establishment , serum replacement , single-cell culture

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Supported by a grant from the Stem Cell Research Center of the 21st Century Frontier Research Program funded by the Ministry of Science and Technology, Gwacheon, Korea (SC-2170). The authors also acknowledge a graduate fellowship provided by the Korean Ministry of Education through the Brain Korea 21 project, Seoul, Korea.

PII: S0015-0282(06)01078-8

doi:10.1016/j.fertnstert.2006.01.056

« Previous Next »Fertility and Sterility
Volume 86, Issue 4, Supplement , Pages 1137-1145, October 2006

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