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Human Long-Term Culture Initiating Cell Assay

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Basic Cell Culture Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 946))

Abstract

The long-term culture initiating cell (LTC-IC) assay, founded on the bone marrow long-term culture (LTC) system, measures primitive hematopoietic stem cells (termed LTC-IC) based on their capacity to produce myeloid progeny for at least 5 weeks. Adaptations of the LTC system including the use of stromal cell lines, application of limiting dilution analysis, and estimation of average hematopoietic progenitor output per LTC-IC under defined conditions have made it possible to accurately determine LTC-IC content in minimally separated and highly purified cell populations from human hematopoietic tissue sources such as bone marrow, peripheral blood, cord blood, fetal liver as well as cord blood and mobilized peripheral blood. Methodologies for measuring human LTC-IC using bulk cultures, limiting dilution analysis, and single cell cultures are described.

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Author information

Authors and Affiliations

  1. Terry Fox Laboratory, Vancouver, BC, Canada

    Min Liu

  2. Private Chinese Culture University, Taipei, Taiwan

    Min Liu

  3. STEMCELL Technologies, Vancouver, BC, Canada

    Cindy L. Miller

  4. Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, BC, Canada

    Connie J. Eaves

Authors
  1. Min Liu
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  2. Cindy L. Miller
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  3. Connie J. Eaves
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Corresponding author

Correspondence to Cindy L. Miller .

Editor information

Editors and Affiliations

  1. , Experimental Therapeutics, BC Cancer Agency, West 10th Ave. 675, Vancouver, V5Z 1L3, British Columbia, Canada

    Cheryl D. Helgason

  2. STEMCELL Technologies, Inc., W. Seventh Avenue 570, Vancouver, V5Z 1B3, British Columbia, Canada

    Cindy L. Miller

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Liu, M., Miller, C.L., Eaves, C.J. (2013). Human Long-Term Culture Initiating Cell Assay. In: Helgason, C., Miller, C. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology, vol 946. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-128-8_15

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  • DOI: https://doi.org/10.1007/978-1-62703-128-8_15

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-127-1

  • Online ISBN: 978-1-62703-128-8

  • eBook Packages: Springer Protocols

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