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* Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, Ghent, Belgium;
Department of Pathology, AZ St-Jan Hospital, Bruges, Belgium;
Centre for Stem Cell Biology, University of Sheffield, Sheffield, United Kingdom; and
Department of Immunology, Erasmus Medical Center, Rotterdam, The Netherlands
Human embryonic stem cells (hESC) are pluripotent stem cells. A major challenge in the field of hESC is the establishment of specific differentiation protocols that drives hESC down a particular lineage fate. So far, attempts to generate T cells from hESC in vitro were unsuccessful. In this study, we show that T cells can be generated in vitro from hESC-derived hematopoietic precursor cells present in hematopoietic zones (HZs). These zones are morphologically similar to blood islands during embryonic development, and are formed when hESC are cultured on OP9 stromal cells. Upon subsequent transfer of these HZs on OP9 cells expressing high levels of Delta-like 1 and in the presence of growth factors, cells expand and differentiate to T cells. Furthermore, we show that T cells derive exclusively from a CD34highCD43low population, further substantiating the notion that hESC-derived CD34highCD43low cells are formed in HZs and are the only population containing multipotent hematopoietic precursor cells. Differentiation to T cells sequentially passes through the physiological intermediates: CD34+CD7+ T/NK committed, CD7+CD4+CD8– immature single positive, CD4+CD8+ double positive, and finally CD3+CD1–CD27+ mature T cell stages. TCR
β+ and TCR
+ T cells are generated. Mature T cells are polyclonal, proliferate, and secrete cytokines in response to mitogens. This protocol for the de novo generation of T cells from hESC could be clinically and scientifically relevant.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 This work was supported by Grant G.0096.05 of the Fund for Scientific Research, Flanders (FWO Vlaanderen) and the Interuniversity Attraction Poles Program (IUAP), supported by the Belgian Science Policy. F.T., T.T., and T.K. are supported by the Fund for Scientific Research, Flanders (FWO Vlaanderen). I.V. is supported by the Interuniversity Attraction Poles Program (IUAP). S.V.C. is supported by the Instituut voor de Aanmoediging van Innovatie door Wetenschap en Technologie in Vlaanderen (IWT).
2 Address correspondence and reprint requests to Bart Vandekerckhove, Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, University Hospital Ghent, Blok A, 4th floor, De Pintelaan 185, Ghent, Belgium. E-mail address: bart.vandekerckhove{at}ugent.be
3 Abbreviations used in this paper: hESC, human embryonic stem cell; HPC, hematopoietic precursor cell; OP9-DL1, cocultures with Delta-like 1 expressing OP9 stromal cell; BM, bone marrow; HZ, hematopoietic zone; iPS cell, induced pluripotent stem cell; SCF, stem cell factor; DP, double positive; HSC, hematopoietic stem cell; cyCD3
, cytoplasmic CD3; VEGFR-2, vascular endothelial growth factor receptor-2.
4 The online version of this article contains supplementary material.
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  D. S. Kaufman Toward clinical therapies using hematopoietic cells derived from human pluripotent stem cells Blood, October 22, 2009; 114(17): 3513 - 3523. [Abstract] [Full Text] [PDF]
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