Long-Term Culture-Initiating Cell Assay for Mouse Cells

  • Stefan Woehrer
  • Cindy L. Miller
  • Connie J. Eaves
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 946)

Abstract

The long-term culture-initiating cell (LTC-IC) assay is a well-established in vitro assay used to enumerate primitive mouse hematopoietic stem cells (HSCs) and relies on the two cardinal functions of HSCs: ability to self-renew and differentiation capacity. LTC-ICs present in minimally processed and purified cell suspensions and cocultured on a supportive feeder layer are detected by their sustained ability to produce hematopoietic progenitors (colony forming cells) after ≥ 4 weeks in culture. Refinements including the use of a defined stromal cell line, and extending the in vitro culture to 6 weeks allow detection of LTC-IC at similar frequencies to transplantable HSCs quantified using in vivo assays.

Key words

Hematopoietic stem cells Stromal cells Stem cell niche LTC-IC Long-term culture-initiating cell 

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Stefan Woehrer
    • 1
    • 2
  • Cindy L. Miller
    • 3
  • Connie J. Eaves
    • 4
  1. 1.Terry Fox LaboratoryBC Cancer AgencyVancouverCanada
  2. 2.Medical University of ViennaViennaAustria
  3. 3.STEMCELL TechnologiesVancouverCanada
  4. 4.Terry Fox LaboratoryBritish Columbia Cancer AgencyVancouverCanada

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