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Revised “hPSC-Sac Method” for Simple and Efficient Differentiation of Human Pluripotent Stem Cells to Hematopoietic Progenitor Cells

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Induced Pluripotent Stem (iPS) Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2454))

Abstract

The human hematopoietic differentiation in vitro of human pluripotent stem cells (hPSCs) has provided new tools to elucidate the mechanisms of related genetic abnormalities, such as congenital diseases and acquired hematopoietic malignancies, and to discover new treatments. The differentiation can also be applied to developing a stable source of blood products for transfusion with minimal risk of several blood-borne infections. We previously proposed a method for hematopoietic progenitor cell (HPC) differentiation, the “hPSC-sac method”, in which hPSCs are cocultured with C3H10T1/2 mouse stromal cells and mixed with a single cytokine, VEGF. The hPSC-sac method can differentiate hPSCs to multiple blood lineages. Here we describe improvements in the method by adding bFGF, TGFβ inhibitor and heparin to the culture, which increases the yield of CD34+CD43+ HPCs 50-fold compared with the original protocol. This revised hPSC-sac method is expected to contribute to the development of disease models and regenerative medicine using hematopoietic lineage cells.

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Acknowledgments

The authors thank Dr. Naoshi Sugimoto and Dr. Peter Karangiannis for critical reading of the manuscript. This work was supported in part by the Highway Program for Realization of Regenerative Medicine (JP18bm0504008, K.E.) and the Core Center for iPS Cell Research (JP18bm0104001, K.E.) from the Japan Agency for Medical Research and Development (AMED), and by Grant-in-Aid for JSPS fellow (JP18J14237, A.Y.) and for scientific research (18H04164, K.E.) from the Japan Society for the Promotion of Science (JSPS).

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Authors and Affiliations

  1. Department of Clinical Application, CiRA, Kyoto University, Kyoto, Japan

    Akinori Yuzuriha & Koji Eto

  2. Department of Regenerative Medicine, Chiba University Graduate School of Medicine, Chiba, Japan

    Koji Eto

Authors
  1. Akinori Yuzuriha
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  2. Koji Eto
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Corresponding author

Correspondence to Koji Eto .

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Editors and Affiliations

  1. Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, ON, Canada

    Andras Nagy

  2. Ottawa, ON, Canada

    Kursad Turksen

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Yuzuriha, A., Eto, K. (2021). Revised “hPSC-Sac Method” for Simple and Efficient Differentiation of Human Pluripotent Stem Cells to Hematopoietic Progenitor Cells. In: Nagy, A., Turksen, K. (eds) Induced Pluripotent Stem (iPS) Cells. Methods in Molecular Biology, vol 2454. Humana, New York, NY. https://doi.org/10.1007/7651_2021_443

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  • DOI: https://doi.org/10.1007/7651_2021_443

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2118-9

  • Online ISBN: 978-1-0716-2119-6

  • eBook Packages: Springer Protocols

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