Abstract
With their unique capabilities of self-renewal and differentiation into three germ layers, mouse embryonic stem cells (mESCs) are widely used as an in vitro cellular model for early mammalian developmental studies. mESCs are traditionally cultured in high-serum and LIF-containing medium on a growth-deficient mouse embryonic fibroblast layer. A more recent culturing system with two inhibitors (for GSK3β (CHIR99021) and MEK1/2 (PD0325901)) and LIF enables the derivation of mESC lines from various mouse strains. Here we describe methods for the mESC growth and maintenance in each medium composition as well as their adaptation to either condition.
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References
Huang G, Ye S, Zhou X et al (2015) Molecular basis of embryonic stem cell self-renewal: from signaling pathways to pluripotency network. Cell Mol Life Sci 72:1741–1757
Niwa H (2014) The pluripotency transcription factor network at work in reprogramming. Curr Opin Genet Dev 28:25–31
Ying QL, Smith A (2017) The art of capturing pluripotency: creating the right culture. Stem Cell Rep 8:1457–1464
Wray J, Kalkan T, Smith AG (2010) The ground state of pluripotency. Biochem Soc Trans 38:1027–1,032
Nichols J, Smith A (2009) Naive and primed pluripotent states. Cell Stem Cell 4:487–492
Tamm C, Galitó SP, Annerén C (2013) A comparative study of protocols for mouse embryonic stem cell culturing. PLoS One 8:1–17
Chaudhry MA, Vitalis TZ, Bowen BD et al (2008) Basal medium composition and serum or serum replacement concentration influences on the maintenance of murine embryonic stem cells. Cytotechnology 58:173–179
Marks H, Kalkan T, Menafra R et al (2012) The transcriptional and epigenomic foundations of ground state pluripotency. Cell 149:590–604
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Balbasi, E., Guven, G., Terzi Cizmecioglu, N. (2021). Mouse Embryonic Stem Cell Culture in Serum-Containing or 2i Conditions. In: Turksen, K. (eds) Embryonic Stem Cell Protocols . Methods in Molecular Biology, vol 2520. Humana, New York, NY. https://doi.org/10.1007/7651_2021_438
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DOI: https://doi.org/10.1007/7651_2021_438
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2436-4
Online ISBN: 978-1-0716-2437-1
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