Abstract
Human embryonic stem cells (hESCs) and patient-specific human induced pluripotent stem cells (hiPSCs) are valuable reagents for studying the earliest stages of hematopoietic genesis and for modeling the developmental basis of hematologic disorders, and they may also have the potential for generating an unlimited supply of autologous transplantable hematopoietic stem cells. We have previously described the development phases of hematopoiesis that arise from differentiated hESCs using a human embryoid body (hEB) culture system. This hEB system produces hematopoietic and endothelial progenitors through a hemangioblast intermediate. In this chapter, we describe a modified and optimized version of this hEB system that generates high frequencies of multipotent CD34+CD45+ hematopoietic progenitors via employing hEB hematopoietic differentiation followed by adherent hemogenic endothelium generation. Multipotent hematopoietic progenitors can be reproducibly generated in robust amounts with this differentiation system from a wide variety of hiPSC or hESC for biochemical, transcriptomic, epigenetic, and transplantation studies.
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References
Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T., Tomoda, K., and Yamanaka, S. (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors, Cell 131, 861–872.
Yu, J., Vodyanik, M. A., Smuga-Otto, K., Antosiewicz-Bourget, J., Frane, J. L., Tian, S., Nie, J., Jonsdottir, G. A., Ruotti, V., Stewart, R., Slukvin, II, and Thomson, J. A. (2007) Induced pluripotent stem cell lines derived from human somatic cells, Science 318, 1917–1920.
Park, I. H., Arora, N., Huo, H., Maherali, N., Ahfeldt, T., Shimamura, A., Lensch, M. W., Cowan, C., Hochedlinger, K., and Daley, G. Q. (2008) Disease-specific induced pluripotent stem cells, Cell 134, 877–886.
Zambidis, E. T., Peault, B., Park, T. S., Bunz, F., and Civin, C. I. (2005) Hematopoietic differentiation of human embryonic stem cells progresses through sequential hematoendothelial, primitive, and definitive stages resembling human yolk sac development, Blood 106, 860–870.
Zambidis, E. T., Park, T. S., Yu, W., Tam, A., Levine, M., Yuan, X., Pryzhkova, M., and Peault, B. (2008) Expression of angiotensin-converting enzyme (CD143) identifies and regulates primitive hemangioblasts derived from human pluripotent stem cells, Blood 112, 3601–3614.
Kim, K., Doi, A., Wen, B., Ng, K., Zhao, R., Cahan, P., Kim, J., Aryee, M. J., Ji, H., Ehrlich, L. I., Yabuuchi, A., Takeuchi, A., Cunniff, K. C., Hongguang, H., McKinney-Freeman, S., Naveiras, O., Yoon, T. J., Irizarry, R. A., Jung, N., Seita, J., Hanna, J., Murakami, P., Jaenisch, R., Weissleder, R., Orkin, S. H., Weissman, I. L., Feinberg, A. P., and Daley, G. Q. (2010) Epigenetic memory in induced pluripotent stem cells, Nature.
Yu, J., Hu, K., Smuga-Otto, K., Tian, S., Stewart, R., Slukvin, I. I., and Thomson, J. A. (2009) Human induced pluripotent stem cells free of vector and transgene sequences. Science 324, 797–801.
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© 2011 Humana Press
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Park, T.S., Burridge, P.W., Zambidis, E.T. (2011). Generation of Multipotent CD34+CD45+ Hematopoietic Progenitors from Human Induced Pluripotent Stem Cells. In: Ye, K., Jin, S. (eds) Human Embryonic and Induced Pluripotent Stem Cells. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-61779-267-0_24
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DOI: https://doi.org/10.1007/978-1-61779-267-0_24
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