Advertisement

Pig Chimeric Model with Human Pluripotent Stem Cells

  • Cuiqing Zhong
  • Jun WuEmail author
  • Juan Carlos Izpisua BelmonteEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2005)

Abstract

Interspecies chimera formation provides a unique platform for studying donor cell developmental potential, modeling disease in vivo, as well as in vivo production of tissues and organs. The derivation of human pluripotent stem cells (hPSC) from either human embryos or somatic cell reprogramming facilitates our understanding of human development, as well as accelerates our exploration of regenerative medicine for human health. Due to similar organ size, close anatomy, and physiology between pig and human, human-Pig interspecies chimeric model in which pig serves as the host species may open new avenues for studying human embryogenesis, disease pathogenesis, and generation of human organ for transplantation to solve the worldwide donor organ shortage. Our previous study demonstrated chimeric competency of different types of human PSCs in pig host. In this chapter, we introduce our workflow for the generation of human PSCs and analysis of its chimeric contribution to pre- and postimplantation pig embryos.

Key words

Human pluripotent stem cells (hPSCs) Human embryonic stem cells (hESCs) Human induced pluripotent stem cells (hiPSCs) Blastocyst Epiblast stem cells (EpiSCs) Pig Chimeric contribution 

Notes

Acknowledgment

We would like to thank Salk Waitt Advanced Biophotonic Core for technical advice on imaging analysis and Salk Stem Cell Core for providing cell culture reagents. We would like to thank May Schwarz and Peter Schwarz for administrative help. This work was supported by Universidad Católica San Antonio de Murcia (UCAM), the Larry L. Hillblom Foundation, Paul F. Glenn Foundation, and the Moxie Foundation.

References

  1. 1.
    Evans MJ, Kaufman MH (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292(5819):154–156CrossRefGoogle Scholar
  2. 2.
    Thomson JA et al (1998) Embryonic stem cell lines derived from human blastocysts. Science 282(5391):1145–1147CrossRefGoogle Scholar
  3. 3.
    Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126(4):663–676CrossRefGoogle Scholar
  4. 4.
    Takahashi K et al (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131(5):861–872CrossRefGoogle Scholar
  5. 5.
    Brons IG et al (2007) Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature 448(7150):191–195CrossRefGoogle Scholar
  6. 6.
    Tesar PJ et al (2007) New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 448(7150):196–199CrossRefGoogle Scholar
  7. 7.
    Nichols J, Smith A (2009) Naive and primed pluripotent states. Cell Stem Cell 4(6):487–492CrossRefGoogle Scholar
  8. 8.
    Wu J et al (2015) An alternative pluripotent state confers interspecies chimaeric competency. Nature 521(7552):316–321CrossRefGoogle Scholar
  9. 9.
    Wu J, Izpisua Belmonte JC (2016) Stem cells: a renaissance in human biology research. Cell 165(7):1572–1585CrossRefGoogle Scholar
  10. 10.
    Ying QL et al (2008) The ground state of embryonic stem cell self-renewal. Nature 453(7194):519–523CrossRefGoogle Scholar
  11. 11.
    Huang Y et al (2012) In Vivo differentiation potential of epiblast stem cells revealed by chimeric embryo formation. Cell Rep 2(6):1571–1578CrossRefGoogle Scholar
  12. 12.
    Gafni O et al (2013) Derivation of novel human ground state naive pluripotent stem cells. Nature 504(7479):282–286CrossRefGoogle Scholar
  13. 13.
    Takashima Y et al (2014) Resetting transcription factor control circuitry toward ground-state pluripotency in human. Cell 158(6):1254–1269CrossRefGoogle Scholar
  14. 14.
    Theunissen TW et al (2014) Systematic identification of culture conditions for induction and maintenance of naive human pluripotency. Cell Stem Cell 15(4):471–487CrossRefGoogle Scholar
  15. 15.
    Wu J, Izpisua Belmonte JC (2015) Dynamic pluripotent stem cell states and their applications. Cell Stem Cell 17(5):509–525CrossRefGoogle Scholar
  16. 16.
    Wu J et al (2017) Interspecies chimerism with mammalian pluripotent stem cells. Cell 168(3):473–486.e15CrossRefGoogle Scholar
  17. 17.
    Tsukiyama T, Ohinata Y (2014) A modified EpiSC culture condition containing a GSK3 inhibitor can support germline-competent pluripotency in mice. PLoS One 9(4):e95329CrossRefGoogle Scholar
  18. 18.
    Yoshioka K, Noguchi M, Suzuki C (2012) Production of piglets from in vitro-produced embryos following non-surgical transfer. Anim Reprod Sci 131(1–2):23–29CrossRefGoogle Scholar
  19. 19.
    Pursel VG, Johnson LA (1975) Freezing of boar spermatozoa: fertilizing capacity with concentrated semen and a new thawing procedure. J Anim Sci 40(1):99–102CrossRefGoogle Scholar
  20. 20.
    Funahashi H, Ekwall H, Rodriguez-Martinez H (2000) Zona reaction in porcine oocytes fertilized in vivo and in vitro as seen with scanning electron microscopy. Biol Reprod 63(5):1437–1442CrossRefGoogle Scholar
  21. 21.
    Martinez EA et al (2014) Successful non-surgical deep uterine transfer of porcine morulae after 24 hour culture in a chemically defined medium. PLoS One 9(8):e104696CrossRefGoogle Scholar
  22. 22.
    Petters RM, Wells KD (1993) Culture of pig embryos. J Reprod Fertil Suppl 48:61–73PubMedGoogle Scholar
  23. 23.
    Ross PJ et al (2008) Polycomb gene expression and histone H3 lysine 27 trimethylation changes during bovine preimplantation development. Reproduction 136(6):777–785CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Salk Institute for Biological StudiesLa JollaUSA
  2. 2.Department of Molecular BiologyUniversity of Texas Southwestern Medical CenterDallasUSA

Personalised recommendations