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Hematopoietic Differentiation of Pluripotent Stem Cells in Culture

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Hematopoietic Stem Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1185))

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Abstract

This chapter describes a two-dimensional “monolayer” system for differentiating human pluripotent stem cells (PSCs) into “primitive” hematopoietic progenitor cells (HPCs) resembling those produced in vivo by the early embryonic yolk sac. This experimental system utilizes defined conditions without serum or feeder cells. Cytokines are added sequentially to stimulate the formation of mesoderm and its subsequent patterning to hematopoietic progenitors. The HPCs produced by this protocol have multi-lineage potential (erythroid, megakaryocyte, and myeloid) and can be isolated as a homogeneous population for use in standard hematopoietic studies including liquid expansion to mature lineages and colony assays. In addition, the HPCs can be cryopreserved for distribution or analysis at later times. The HPCs generated by this protocol have been used successfully to better define intrinsic variation in hematopoietic potential between different PSC lines and to model human hematopoietic diseases using patient-derived induced pluripotent stem cells.

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Author information

Author notes

    Authors and Affiliations

    1. Department of Pathology and Laboratory Medicine and Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, PA, USA

      Jason A. Mills

    2. Department of Pathology and Laboratory Medicine and Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Colket Translational Research Building 5014, 3501 Civic Center Blvd., Philadelphia, PA, 19104, USA

      Prasuna Paluru, Paul Gadue & Deborah L. French

    3. Department of Pediatrics, The Children’s Hospital of Philadelphia, Philadelphia, PA, USA

      Mitchell J. Weiss

    Authors
    1. Jason A. Mills
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    2. Prasuna Paluru
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    3. Mitchell J. Weiss
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    4. Paul Gadue
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    5. Deborah L. French
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    Corresponding author

    Correspondence to Deborah L. French .

    Editor information

    Editors and Affiliations

    1. Department of Pediatrics, Aflac Cancer and Blood Disorders Center Emory University School of Medicine, Atlanta, Georgia, USA

      Kevin D. Bunting

    2. Department of Pediatrics, Emory University School of Medicine Aflac Cancer & Blood Disorders Ctr, Atlanta, Georgia, USA

      Cheng-Kui Qu

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    © 2014 Springer Science+Business Media New York

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    Mills, J.A., Paluru, P., Weiss, M.J., Gadue, P., French, D.L. (2014). Hematopoietic Differentiation of Pluripotent Stem Cells in Culture. In: Bunting, K., Qu, CK. (eds) Hematopoietic Stem Cell Protocols. Methods in Molecular Biology, vol 1185. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1133-2_12

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    • DOI: https://doi.org/10.1007/978-1-4939-1133-2_12

    • Published:

    • Publisher Name: Humana Press, New York, NY

    • Print ISBN: 978-1-4939-1132-5

    • Online ISBN: 978-1-4939-1133-2

    • eBook Packages: Springer Protocols

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