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Stem Cell Reports. 2014 Jan 30;2(2):232-42. doi: 10.1016/j.stemcr.2013.12.013. eCollection 2014 Feb 11.

Derivation and maintenance of murine trophoblast stem cells under defined conditions.

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Department of Developmental Pathology, Institute of Pathology, University of Bonn, 53127 Bonn, Germany.
Epigenetics Programme, The Babraham Institute, Babraham Research Campus, Cambridge CB22 3AT, UK ; Centre for Trophoblast Research, University of Cambridge, Cambridge CB2 3EG, UK.
Max Planck Institute for Molecular Biomedicine, Group Laboratory of Computational Biology and Bioinformatics, 48149 Münster, Germany.
Life and Brain Center, Institute of Molecular Psychiatry, University of Bonn, 53127 Bonn, Germany.
Life and Brain Center, Institute of Reconstructive Neurobiology, University of Bonn and Hertie Foundation, 53127 Bonn, Germany.


Trophoblast stem cells (TSCs) are in vitro equivalents to the precursor cells of the placenta. TSCs are cultured in serum-rich medium with fibroblast growth factor 4, heparin, and embryonic-fibroblast-conditioned medium. Here, we developed a simple medium consisting of ten chemically defined ingredients for culture of TSCs on Matrigel or synthetic substrates, named TX medium. Gene expression and DNA methylation profiling demonstrated the faithful propagation of expression profiles and epigenomic characteristics of TSCs cultured in TX. Further, TX medium supported the de novo derivation of TSC lines. Finally, TSCs cultured in TX differentiate into all derivatives of the trophectodermal lineage in vitro, give rise to hemorrhagic lesions in nude mice, and chimerize the placenta, indicating that they retained all hallmarks of TSCs. TX media formulation no longer requires fetal bovine serum and conditioned medium, which facilitates and standardizes the culture of this extraembryonic lineage.

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