Skip to main content

Hematopoietic and Nature Killer Cell Development from Human Pluripotent Stem Cells

  • Protocol
  • First Online:
Embryonic Stem Cell Immunobiology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1029))

Abstract

Natural killer (NK) cells are key effectors of the innate immune system, protecting the host from a variety of infections, as well as malignant cells. Recent advances in the field of NK cell biology have led to a better understanding of how NK cells develop. This progress has directly translated to improved outcomes in patients receiving hematopoietic stem cell transplants to treat potentially lethal malignancies. However, key differences between mouse and human NK cell development and biology limits the use of rodents to attain a more in depth understanding of NK cell development. Therefore, a readily accessible and genetically tractable cell source to study human NK cell development is warranted. Our lab has pioneered the development of lymphocytes, specifically NK cells, from human embryonic stem cells (hESCs) and more recently induced pluripotent stem cells (iPSCs). This chapter describes a reliable method to generate NK cells from hESCs and iPSCs using murine stromal cell lines. Additionally, we include an updated approach using a spin-embryoid body (spin-EB) differentiation system that allows for human NK cell development completely defined in vitro conditions.

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. Woll PS, Martin CH, Miller JS, Kaufman DS (2005) Human embryonic stem cell-derived NK cells acquire functional receptors and cytolytic activity. J Immunol 175:5095–5103

    PubMed  CAS  Google Scholar 

  2. Woll PS, Grzywacz B, Tian X et al (2009) Human embryonic stem cells differentiate into a homogeneous population of natural killer cells with potent in vivo antitumor activity. Blood 113:6094–6101

    Article  PubMed  CAS  Google Scholar 

  3. Ni Z, Knorr DA, Clouser CL et al (2011) Human pluripotent stem cells produce natural killer cells that mediate anti-HIV-1 activity utilizing diverse cellular mechanisms. J Virol 85(1):43–50

    Article  PubMed  CAS  Google Scholar 

  4. Wilber A, Linehan JL, Tian X et al (2007) Efficient and stable transgene expression in human embryonic stem cells using transposon-­mediated gene transfer (Dayton, Ohio). Stem Cells 25:2919–2927

    Article  PubMed  CAS  Google Scholar 

  5. Giudice A, Trounson A (2008) Genetic modification of human embryonic stem cells for derivation of target cells. Cell Stem Cell 2:422–433

    Article  PubMed  CAS  Google Scholar 

  6. Ng ES, Davis R, Stanley EG, Elefanty AG (2008) A protocol describing the use of a recombinant protein-based, animal product-­free medium (APEL) for human embryonic stem cell differentiation as spin embryoid bodies. Nat Protoc 3:768–776

    Article  PubMed  CAS  Google Scholar 

  7. Ng ES, Davis RP, Azzola L, Stanley EG, Elefanty AG (2005) Forced aggregation of defined numbers of human embryonic stem cells into embryoid bodies fosters robust, reproducible hematopoietic differentiation. Blood 106:1601–1603

    Article  PubMed  CAS  Google Scholar 

  8. Kaufman DS, Hanson ET, Lewis RL, Auerbach R, Thomson JA (2001) Hematopoietic colony-­forming cells derived from human embryonic stem cells. Proc Natl Acad Sci U S A 98:10716–10721

    Article  PubMed  CAS  Google Scholar 

  9. Ledran MH, Krassowska A, Armstrong L et al (2008) Efficient hematopoietic differentiation of human embryonic stem cells on stromal cells derived from hematopoietic niches. Cell Stem Cell 3:85–98

    Article  PubMed  CAS  Google Scholar 

  10. Tian X, Morris JK, Linehan JL, Kaufman DS (2004) Cytokine requirements differ for stroma and embryoid body-mediated hematopoiesis from human embryonic stem cells. Exp Hematol 32:1000–1009

    Article  PubMed  CAS  Google Scholar 

  11. Ng ES, Davis RP, Hatzistavrou T, Stanley EG, Elefanty AG (2008) Directed differentiation of human embryonic stem cells as spin embryoid bodies and a description of the hematopoietic blast colony forming assay. Curr Protoc Stem Cell Biol; Chapter 1:Unit 1D 3

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media New York

About this protocol

Cite this protocol

Ni, Z., Knorr, D.A., Kaufman, D.S. (2013). Hematopoietic and Nature Killer Cell Development from Human Pluripotent Stem Cells. In: Zavazava, N. (eds) Embryonic Stem Cell Immunobiology. Methods in Molecular Biology, vol 1029. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-478-4_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-478-4_3

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-477-7

  • Online ISBN: 978-1-62703-478-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics