Abstract
In the mouse preimplantation embryo, the first cell fate determination segregates two morphologically and functionally distinct cell lineages. One is the inner cell mass, and the other is the trophectoderm. A subset of the trophectoderm maintains a proliferative capacity and forms the extraembryonic ectoderm, the ectoplacental cone, and the secondary giant cells of the early conceptus after implantation. A stem cell population of the trophectoderm lineage can be isolated and maintained in vitro under the presence of fibroblast growth factor 4, heparin, and a feeder layer of mouse embryonic fibroblast cells. Such apparently immortal stem cells, trophoblast stem (TS) cells, exhibit the potential to differentiate to multiple cell types in vitro. TS cells also have the ability to contribute to normal development in chimeras. However, TS cells exclusively contribute to the trophoblastic component of the placenta and of the parietal yolk sac, making a striking contrast with embryonic stem cells, which never contribute to these tissues in chimeras. In this chapter, detailed protocols for the isolation and establishment of TS cell lines from blastocysts and their maintenance are described.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Rossant J. and Cross J. C. (2001) Placental development: lessons from mouse mutants. Nat. Rev. Genet. 2, 538–548.
Tanaka S., Kunath T., Hadjantonakis A. K., Nagy A., and Rossant J. (1998) Promotion of trophoblast stem cell proliferation by FGF4. Science 282, 2072–2075.
Yan J., Tanaka S., Oda M., Makino T., Ohgane J., and Shiota K. (2001) Retinoic acid promotes differentiation of trophoblast stem cells to a giant cell fate. Dev. Biol. 235, 422–432.
Hughes M., Dobric N., Scott I. C., et al. (2004) The Hand1, Stra13 and Gcm1 transcription factors override FGF signaling to promote terminal differentiation of trophoblast stem cells. Dev. Biol. 271, 26–37.
Kunath T., Strumpf D., and Rossant J. (2004) Early trophoblast determination and stem cell maintenance in the mouse-a review. Placenta 25,Suppl. A, S32–S38.
Tanaka T. S., Kunath T., Kimber W. L., et al. (2004) Gene expression profiling of embryoderived stem cells reveals candidate genes associated with pluripotency and lineage specificity. Genome Res. 12, 1921–1928.
Sharov A. A., Piao Y., Matoba R., et al. (2003) Transcriptome analysis of mouse stem cells and early embryos. PLoS Biol. 1, 410–419.
Shiota K., Kogo Y., Ohgane J., et al. (2004) Epigenetic marks by DNA methylation specific to stem, germ and somatic cells in mice. Genes Cells 7, 961–969.
Uy G. D., Downs K. M., and Gardner R. L. (2002) Inhibition of trophoblast stem cell potential in chorionic ectoderm coincides with occlusion of the ectoplacental cavity in the mouse. Development 129, 3913–3924.
Nagy A., Gertsenstein M., Vintersten K., and Behringer R. (2003) Manipulating the Mouse Embryo, 3rd ed., Cold Spring Harbor Laboratory Press, New York.
Kunath T., Strumpf D., Rossant J., and Tanaka S. (2001) Trophoblast stem cells, in Stem Cell Biology (Marshak D. R., Gardner R. L., and Gottlieb D., eds.), Cold Spring Harbor Laboratory Press, New York, pp. 267–286.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Tanaka, S. (2006). Derivation and Culture of Mouse Trophoblast Stem Cells In Vitro. In: Turksen, K. (eds) Embryonic Stem Cell Protocols. Methods in Molecular Biology, vol 329. Humana Press. https://doi.org/10.1385/1-59745-037-5:35
Download citation
DOI: https://doi.org/10.1385/1-59745-037-5:35
Publisher Name: Humana Press
Print ISBN: 978-1-58829-498-2
Online ISBN: 978-1-59745-037-9
eBook Packages: Springer Protocols