Abstract
Embryonic stem (ES) cells have been routinely used to create loss-of-function mutations or gene replacement by homologous recombination in mice, providing an invaluable tool to address fundamental biological questions. Although mouse ES cells have been available for the past 29 years, authentic rat ES cells have only recently been established. The efficient derivation of multiple rat ES cell lines by independent investigators will accelerate the development of novel laboratory tools for biomedical research. Here we provide detailed protocols for the derivation and propagation of rat ES cells and for the production of rat chimeras using rat ES cells. The availability of rat ES cells provides the opportunity to adapt the technology developed in the mouse to the rat.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Abbreviations
- BMP:
-
Bone morphogenetic protein
- DA rat:
-
Dark Agouti rat
- Dpc:
-
Day post-coitum
- ES cells:
-
Embryonic stem cells
- F344 rat:
-
Fischer 344 rat
- GSK3:
-
Glycogen synthase kinase 3
- ICM:
-
Inner cell mass
- IP:
-
Intraperitoneal
- LIF:
-
Leukemia inhibitory factor
- MAPK:
-
Mitogen-activated protein kinase
- MEFs:
-
Mouse embryonic fibroblasts
- PBS:
-
Phosphate-buffered saline
- SD rat:
-
Sprague Dawley rat
References
Aitman TJ, Critser JK, Cuppen E, Dominiczak A, Fernandez-Suarez XM, Flint J, Gauguier D, Geurts AM, Gould M, Harris PC et al (2008) Progress and prospects in rat genetics: a community view. Nat Genet 40:516–522
Evans MJ, Kaufman MH (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292:154–156
Martin GR (1981) Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci USA 78:7634–7638
Buehr M, Meek S, Blair K, Yang J, Ure J, Silva J, McLay R, Hall J, Ying QL, Smith A (2008) Capture of authentic embryonic stem cells from rat blastocysts. Cell 135:1287–1298
Li P, Tong C, Mehrian-Shai R, Jia L, Wu N, Yan Y, Maxson RE, Schulze EN, Song H, Hsieh CL et al (2008) Germline competent embryonic stem cells derived from rat blastocysts. Cell 135:1299–1310
Smith AG, Heath JK, Donaldson DD, Wong GG, Moreau J, Stahl M, Rogers D (1988) Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides. Nature 336:688–690
Williams RL, Hilton DJ, Pease S, Willson TA, Stewart CL, Gearing DP, Wagner EF, Metcalf D, Nicola NA, Gough NM (1988) Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature 336:684–687
Ying QL, Nichols J, Chambers I, Smith A (2003) BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell 115:281–292
Ying QL, Wray J, Nichols J, Batlle-Morera L, Doble B, Woodgett J, Cohen P, Smith A (2008) The ground state of embryonic stem cell self-renewal. Nature 453:519–523
Brook FA, Gardner RL (1997) The origin and efficient derivation of embryonic stem cells in the mouse. Proc Natl Acad Sci USA 94:5709–5712
Keller G (2005) Embryonic stem cell differentiation: emergence of a new era in biology and medicine. Genes Dev 19:1129–1155
Capecchi MR (2005) Gene targeting in mice: functional analysis of the mammalian genome for the twenty-first century. Nat Rev Genet 6:507–512
Nagy A, Gertsenstein M, Vintersten K, Behringer R (2003) Manipulating the mouse embryo, a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, New York
Schwartzberg PL, Goff SP, Robertson EJ (1989) Germ-line transmission of a c-abl mutation produced by targeted gene disruption in ES cells. Science 246:799–803
Seong E, Saunders TL, Stewart CL, Burmeister M (2004) To knockout in 129 or in C57BL/6: that is the question. Trends Genet 20:59–62
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer-Verlag Berlin Heidelberg
About this protocol
Cite this protocol
Li, P., Schulze, E.N., Tong, C., Ying, QL. (2011). Rat Embryonic Stem Cell Derivation and Propagation. In: Pease, S., Saunders, T. (eds) Advanced Protocols for Animal Transgenesis. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-20792-1_19
Download citation
DOI: https://doi.org/10.1007/978-3-642-20792-1_19
Published:
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-20791-4
Online ISBN: 978-3-642-20792-1
eBook Packages: Springer Protocols