Abstract
Embryonic stem (ES) cells have been used extensively for site-specific gene targeting in the mouse. The resulting knock-out and knock-in mouse models generated so far have demonstrated their usefulness in biomedical research. However, for many diseases and fields of study, the rat still represents a superior model. The derivation and culture of germline-competent ES cells in the rat would allow the application of site-specific gene targeting technologies to this species of indisputable importance to biomedical research. We have recently shown the derivation, culture, and for the first time, in vivo contribution of rat ES-like cells to developing tissues. This represents an important step forward in making gene targeting technologies available to the rat research community, via development of rat ES cells. Here, we describe the materials, methods and techniques that have been used to obtain rat blastocysts, derive and culture embryonic cell lines from these, and assess the developmental capacity of the cells in vivo.
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References
Aitman TJ, Critser JK, Cuppen E, Dominiczak A, Fernandez-Suarez XM, Flint J et al (2008) Progress and prospects in rat genetics: a community view. Nat Genet 40:516–522
Tesson L, Cozzi J, Menoret S, Remy S, Usal C, Fraichard A et al (2005) Transgenic modifications of the rat genome. Transgenic Res 14:531–546
Ueda S, Kawamata M, Teratani T, Shimizu T, Tamai Y, Ogawa H et al (2008) Establishment of rat embryonic stem cells and making of chimera rats. PLoS ONE 3:e2800
Demers SP, Yoo JG, Lian L, Therrien J, Smith LC (2007) Rat Embryonic Stem-Like (ES-Like) Cells Can Contribute to Extraembryonic Tissues In Vivo. Cloning Stem Cells 9:512–522
Hogan B (1994) Manipulating the mouse embryo : a laboratory manual. Cold Spring Harbor Laboratory Press, Plainview, N.Y
Nagy A (2003) Manipulating the mouse embryo : a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y
Hoffman LM, Carpenter MK (2005) Characterization and culture of human embryonic stem cells. Nat Biotechnol 23:699–708
Van der Auwera I, D’Hooghe T (2001) Superovulation of female mice delays embryonic and fetal development. Hum Reprod 16:1237–1243
Dang ZC, Lowik CW (2005) Removal of serum factors by charcoal treatment promotes adipogenesis via a MAPK-dependent pathway. Mol Cell Biochem 268:159–167
Acknowledgments
The work described here was funded by the Natural Sciences and Engineering Research Council (NSERC) of Canada, (Industrial Post-Graduate Scholarship to S.-P. D.) and by Clonagen inc. The authors thank Carmen Léveillée and Joëlle A. Desmarais for assistance in preparation of this manuscript.
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Demers, SP., Smith, L.C. (2010). Derivation, Culture, and In vivo Developmental Capacity of Embryonic Cell Lines from Rat Blastocysts. In: Anegon, I. (eds) Rat Genomics. Methods in Molecular Biology, vol 597. Humana Press. https://doi.org/10.1007/978-1-60327-389-3_13
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DOI: https://doi.org/10.1007/978-1-60327-389-3_13
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