Skip to main content

Derivation of Human Embryonic Stem Cell Lines from Vitrified Human Blastocysts

  • Protocol
  • First Online:
Human Embryonic Stem Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1307))

Abstract

Human embryonic stem cells are pluripotent cells typically derived from blastulating embryos that have become excess to clinical needs in assisted reproduction programs. They provide cellular models for embryonic development and disease, and are thought to be useful for future cell replacement therapies and regenerative medicine. Here we describe methods to derive human embryonic stem cell lines. This includes blastocyst cryopreservation using a highly efficient vitrification protocol, the production and use of fibroblast feeder cells, embryo plating and passaging of resulting cellular outgrowths, and cryopreservation of putative stem cells lines.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Thomson JA, Itskovitz-Eldor J, Shapiro SS et al (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145–1147

    Article  CAS  PubMed  Google Scholar 

  2. Pouton CW, Haynes JM (2007) Embryonic stem cells as a source of models for drug discovery. Nat Rev Drug Discov 6:605–616

    Article  CAS  PubMed  Google Scholar 

  3. Chong JJ, Yang X, Don CW et al (2014) Human embryonic-stem-cell-derived cardiomyocytes regenerate non-human primate hearts. Nature. doi:10.1038/nature13233

  4. Roy TK, Bradley CK, Bowman MC et al (2014) Single-embryo transfer of vitrified-warmed blastocysts yields equivalent live-birth rates and improved neonatal outcomes compared with fresh transfers. Fertil Steril 101:1294–1301

    Article  PubMed  Google Scholar 

  5. Peura TT, Schaft J, Stojanov T (2010) Derivation of human embryonic stem cell lines from vitrified human embryos. Methods Mol Biol 584:21–54

    Article  PubMed  Google Scholar 

  6. Thomson JA, Kalishman J, Golos TG et al (1995) Isolation of a primate embryonic stem cell line. Proc Natl Acad Sci U S A 92:7844–7848

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Crook JM, Peura TT, Kravets L et al (2007) The generation of six clinical-grade human embryonic stem cell lines. Cell Stem Cell 1:490–494

    Article  CAS  PubMed  Google Scholar 

  8. Peura T, Bosman A, Chami O et al (2008) Karyotypically normal and abnormal human embryonic stem cell lines derived from PGD-analyzed embryos. Cloning Stem Cells 10:203–216

    Article  CAS  PubMed  Google Scholar 

  9. Bradley CK, Scott HA, Chami O et al (2011) Derivation of Huntington’s disease-affected human embryonic stem cell lines. Stem Cells Dev 20:495–502

    Article  CAS  PubMed  Google Scholar 

  10. Bradley CK, Peura T, Dumevska B et al (2014) Cell lines from morphologically abnormal discarded IVF embryos are typically euploid and unaccompanied by intrachromosomal aberrations. Reprod Biomed Online 28:780–788

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Teija T. Peura .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Bradley, C.K., Schaft, J., Roy, T.K., Dumevska, B., Peura, T.T. (2014). Derivation of Human Embryonic Stem Cell Lines from Vitrified Human Blastocysts. In: Turksen, K. (eds) Human Embryonic Stem Cell Protocols. Methods in Molecular Biology, vol 1307. Humana Press, New York, NY. https://doi.org/10.1007/7651_2014_85

Download citation

  • DOI: https://doi.org/10.1007/7651_2014_85

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2667-1

  • Online ISBN: 978-1-4939-2668-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics