Abstract
Pluripotency refers to the potential of cells to generate all cell types of the embryo proper. Pluripotency spans a spectrum of cellular states. At one polar extreme is naïve pluripotency, which is identified based on the potential to form germline chimeras. At the other polar extreme is primed pluripotency, in which pluripotent cells are primed to differentiate. Mouse naïve PS cells can be derived from preimplantation embryos. Primed epiblast stem (EpiS) cells are typically isolated from epiblasts of early postimplantation mouse embryos. In this chapter, we describe protocols for highly efficient derivation and propagation of murine naïve and primed PS cell lines in serum-free conditions from preimplantation and postimplantation embryos. We describe generation of mouse naïve PS cells using LIF and inhibitors of MEK and GSK3 kinases and of mouse primed PS cells using FGF2 and IWR1 compound which induces the stabilization of Axin proteins.
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De Los Angeles A, Ferrari F, Xi R, Fujiwara Y, Benvenisty N, Deng H, Hochedlinger K, Jaenisch R, Lee S, Leitch HG, Lensch MW, Lujan E, Pei D, Rossant J, Wernig M, Park PJ, Daley GQ (2015) Hallmarks of pluripotency. Nature 525(7570):469–478
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. PNAS 78:7634–7638
Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM (1998) Embryonic stem cells derived from human blastocysts. Science 282:1145–1147
Reubinoff BE, Pera MF, Fong CY, Trounson A, Bongso A (2000) Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat Biotechnol 18:399–404
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
Vallier L, Alexander M, Pedersen RA (2005) Activin/Nodal and FGF pathways cooperate to maintain pluripotency of human embryonic stem cells. J Cell Sci 118:4495–4509
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
Wu J, Belmonte JCI (2016) Stem cells: a renaissance in human biology research. Cell 165:1572–1585
Tesar PJ, Chenoweth JG, Brook FA, Davies TJ, Evans EP, Mack DL, Gardner RL, McKay RD (2007) New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 448:196–199
Brons IG, Smithers LE, Trotter MW, Rugg-Gunn P, Sun B, Chuva de Sousa Lopes SM, Howlett SK, Clarkson A, Ahrlund-Richter L, Pedersen RA, Vallier L (2007) Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature 448:191–195
Nichols J, Smith A (2009) Naïve and primed pluripotent states. Cell Stem Cell 4:487–492
Wu J, Izpisua Belmonte JC (2015) Dynamic pluripotent stem cell states and their applications. Cell Stem Cell 17:509–525
Wu J, Okamura D, Li M, Suzuki K, Luo C, Ma L, He Y, Li Z, Benner C, Tamura I, Krause MN, Nery JR, Du T, Zhang Z, Hishida T, Takahashi Y, Aizawa E, Kim NY, Lajara J, Guillen P, Campistol JM, Esteban CR, Ross PJ, Saghatelian A, Ren B, Ecker JR, Izpisua Belmonte JC (2015) An alternative pluripotent state confers interspecies chimeric competency. Nature 521:316–321
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De Los Angeles, A., Okamura, D., Wu, J. (2019). Highly Efficient Derivation of Pluripotent Stem Cells from Mouse Preimplantation and Postimplantation Embryos in Serum-Free Conditions. In: Hyun, I., De Los Angeles, A. (eds) Chimera Research . Methods in Molecular Biology, vol 2005. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9524-0_2
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DOI: https://doi.org/10.1007/978-1-4939-9524-0_2
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