Skip to main content
SpringerLink
Log in
Book cover

Human Pluripotent Stem Cells pp 177–190Cite as

Classical Cytogenetics: Karyotyping Techniques

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 767))

Abstract

Classical cytogenetics by karyotyping has been utilized in clinical research laboratories for more than 50 years and remains the key method used in the stem cell laboratory to assess the genetic stability of stem cell cultures. It is currently the most readily accessible method for detecting chromosomal abnormalities in pluripotent stem cell cultures. This chapter will describe (1) how to prepare a culture to maximize the number of metaphase cells, (2) how to prepare slides containing chromosome spreads (3) methods used to stain chromosomes, and (4) how to interpret the cytogenetic report.

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

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
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

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Thompson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM (1998). Embryonic stem cell lines derived from human blastocysts. Science 282: 1145–1147.

    Article  Google Scholar 

  2. Reubinoff BE, Pera MF, Fong C-Y, Trounson A, Bongso A (2000). Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat Biotechnol 18: 399–404.

    Article  PubMed  CAS  Google Scholar 

  3. Rosler ES, Fisk GJ, Ares X, Irving J, Miura J, Rao MS, Carpenter MK (2003). Long-term culture of human embryonic stem cells in feeder-free conditions. Devl Dyn 229: 259–274.

    Article  Google Scholar 

  4. Holmquist GP and Ashley T (2006). Chromosome organization and chromatin modifications influence gene function and evolution. Cytogenetic Research 114: 96–125

    Article  CAS  Google Scholar 

  5. Yang S, Lin G, Tan YQ, Zhou D, Deng LY, Cheng DH, Lou SW, Liu TC, Zhou XY Sun Z, Xiang Y, Chen TJ, Wen JF, Lu GX (2008). Tumor progression of culture-adapted human embryonic stem cells during long-term culture. Genes Chromosomes Cancer 47: 665–679.

    Article  PubMed  CAS  Google Scholar 

  6. Baker DE, Harrison NJ, Maltby E, Smith K, Moore HD, Shaw PJ, Heath PR, Holden H, Andrews PW (2007). Adaptation to culture of human embryonic stem cells and oncogenesis in vivo. Nat Biotechnol. 25: 207–215.

    Article  PubMed  CAS  Google Scholar 

  7. Draper JS, Smith K, Gokhale P, Moore HD, Maltby E, Johnson J, Meisner L, Zwaka TP, Thomson JA, Adrews PW. (2004) Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nat Biotechnol. 22: 53–54.

    Article  PubMed  CAS  Google Scholar 

  8. Buzzard JJ, Gough, NM, Crook J, Colman A. (2004) Karyotype of human ES cells during extended culture. Nat Biotechnol. 22: 381–382.

    Article  PubMed  CAS  Google Scholar 

  9. Peterson S, Rehen S, Westra W, Yung Y, Chun J (2007) Spectral karyotyping and fluorescent in situ hybridization. In Human Stem Cell Manual: A Laboratory Guide. Loring JF, Wesselschmidt RW, Schwartz PH (eds.). New York: Elsevier.

    Google Scholar 

  10. Gunderson K, Barker DL, Bibikova M, Fan JB (2007) Genotype and epigenotype by single nucleotide polymorphism (SNP) analysis. In Human Stem Cell Manual: A Laboratory Guide. Loring JF, Wesselschmidt RW, Schwartz PH (eds.). New York: Elsevier.

    Google Scholar 

  11. Wesselschmidt RL and Loring JF. (2007) Classical Cytogenetics: Karyotyping. In Human Stem Cell Manual: A Laboratory Guide. Loring JF, Wesselschmidt RW, Schwartz PH (eds.). New York: Elsevier.

    Google Scholar 

  12. Barch, M.J. (ed.) (1991) The ACT Cytogenetics Laboratory Manual, (2nd Edition), Raven Press, New York.

    Google Scholar 

  13. Hsieh, CL. (1998) Basic cytogenetic techniques: culturing, slide making, and G-banding. In Cell Biology: A Laboratory Handbook, (2nd edition.) Vol.3. Elsevier, Academic Press. New York.

    Google Scholar 

  14. ISCN (2005) “An International System for Human Cytogenetic Nomenclature”. S. Karger, Basel/New York.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biology, Beckman Research Institute, City of Hope National Medical Center, Duarte, CA, USA

    Steven E. Bates

Authors
  1. Steven E. Bates
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Steven E. Bates .

Editor information

Editors and Affiliations

  1. Research Institute, CHOC Human Neural Stem Cell Resource, Children's Hospital of Orange County, S. Main Street 455, Orange, 92868-3874, California, USA

    Philip H. Schwartz

  2. Beckman Research Institute, Center for Applied Technology Developmen, City of Hope, East Duarte Road 1500, Duarte, 91010, California, USA

    Robin L. Wesselschmidt

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Bates, S.E. (2011). Classical Cytogenetics: Karyotyping Techniques. In: Schwartz, P., Wesselschmidt, R. (eds) Human Pluripotent Stem Cells. Methods in Molecular Biology, vol 767. Humana Press. https://doi.org/10.1007/978-1-61779-201-4_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-201-4_13

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-200-7

  • Online ISBN: 978-1-61779-201-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation