Abstract
Murine embryonic stem (ES) cells are derived from the inner cell mass of 3.5–day-old embryo and have the ability to colonize the germline and form normal gametes following in vitro genetic manipulations. This remarkable characteristic of ES cells has provided the basis for studying normal gene function in the mouse by targeted mutagenesis. Nevertheless, ES cells are very sensitive and need to be manipulated with care for them to retain totipotency after extensive in vitro manipulations. Here we provide straightforward protocols for proper care of these cells. Special emphasis is placed on aspects that are particularly critical for proper culture of this cell type.
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Acknowledgments
We thank Mary Ellen Palko for critical reading of the manuscript. This research was supported by the Intramural Research Program of the NIH, National Cancer Institute. The contents of this publication do not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. The Center for Cancer Research, NCI-Frederick has filed an Animal Welfare Assurance with the Office for Protection from Research Risks (OPRR). The protocols herein described have been approved by the NCI-Frederick Institutional Animal Care and Use Committee.
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Southon, E., Tessarollo, L. (2009). Manipulating Mouse Embryonic Stem Cells. In: Wurst, W., Kühn, R. (eds) Gene Knockout Protocols. Methods in Molecular Biology, vol 530. Humana Press. https://doi.org/10.1007/978-1-59745-471-1_9
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DOI: https://doi.org/10.1007/978-1-59745-471-1_9
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