Abstract
Mouse embryonic stem cells (mESCs) derived from inner cell mass (ICM) of pre-implantation embryos, can maintain undifferentiated state when cultured in N2B27 medium supplemented with GSK3 inhibitor CHIR99021 and MEK inhibitor PD0325901 ("2i") and leukemia inhibitor factor (LIF). Compare to conventional culture medium, all components of this medium are defined. With the N2B27 medium, “2i” and LIF, mESCs can contribute to the germline of the chimeric embryos, however, whether the “all-ES cells” mice can been generated by tetraploid complementation is unclear yet, while the tetraploid complementation serve as a golden standard to assess the pluripotency of ES cells. Here, our study showed that mESCs derived and cultured with the N2B27 complete medium could generate fertile mice by tetraploid complementation. In addition, the survival rate of tetraploid complementation mice produced by inbred mES cell lines is higher than the conventional culture condition, and increased the percentage of Oct4 positive cells contrast to conventional medium either. Therefore, the N2B27 medium supplemented with “2i” and LIF is an alternative choice for the derivation and long-term culture of mouse embryonic stem cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Evans MJ, Kaufman MH (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292:154–156
Guo XX, He FH (2000) Properties and applications of embryonic stem cells. Chin Sci Bull 45:1258–1265
Smith AG, Heath JK, Donaldson DD et al (1988) Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides. Nature 336:688–690
Niwa H, Burdon T, Chambers I et al (1998) Self-renewal of pluripotent embryonic stem cells is mediated via activation of stat3. Gene Dev 12:2048–2060
Ying QL, Nichols J, Chambers I et al (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, Smith AG (2003) Defined conditions for neural commitment and differentiation. Method Enzymol 365:327–341
Nichols J, Jones K, Phillips JM et al (2009) Validated germline-competent embryonic stem cell lines from nonobese diabetic mice. Nat Med 15:814–818
Shuai L, Feng CJ, Zhang HJ et al (2013) Derivation of androgenetic embryonic stem cells from m-carboxycinnamic acid bishydroxamide (cbha) treated androgenetic embryos. Chin Sci Bull 58:2862–2868
Jiang MG, Li TD, Feng CJ et al (2013) Generation of transgenic rats through induced pluripotent stem cells. J Biol Chem 288:27150–27158
Zhao XY, Li W, Lv Z et al (2009) Ips cells produce viable mice through tetraploid complementation. Nature 461:86–88
Zhao XY, Li W, Lv Z et al (2010) Viable fertile mice generated from fully pluripotent ips cells derived from adult somatic cells. Stem Cell Rev 6:390–397
Wobus AM, Boheler KR (2005) Embryonic stem cells: prospects for developmental biology and cell therapy. Physiol Rev 85:635–678
Wray J, Kalkan T, Smith AG (2010) The ground state of pluripotency. Biochem Soc Trans 38:1027–1032
Smith AG (2001) Embryo-derived stem cells: of mice and men. Annu Rev Cell Dev Biol 17:435–462
Dean W, Bowden L, Aitchison A et al (1998) Altered imprinted gene methylation and expression in completely es cell-derived mouse fetuses: association with aberrant phenotypes. Development 125:2273–2282
Acknowledgements
This work was supported by the National Basic Research Program of China (2012CBA01300 and 2012CB966500).
Author information
Authors and Affiliations
Corresponding author
Additional information
Chunjing Feng and Haifeng Wan contributed equally to this work.
About this article
Cite this article
Feng, C., Wan, H., Zhao, XY. et al. Generation of tetraploid complementation mice from embryonic stem cells cultured with chemical defined medium. Chin. Sci. Bull. 59, 2743–2748 (2014). https://doi.org/10.1007/s11434-014-0395-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11434-014-0395-2