Engineering the Human Thymic Microenvironment to Support Thymopoiesis In Vivo

Authors

  • Brile Chung,

    1. Department of Pathology and Laboratory Medicine, University of California, Los Angeles (UCLA), California, USA
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  • Amélie Montel-Hagen,

    1. Department of Pathology and Laboratory Medicine, University of California, Los Angeles (UCLA), California, USA
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  • Shundi Ge,

    1. Department of Pathology and Laboratory Medicine, University of California, Los Angeles (UCLA), California, USA
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  • Garrett Blumberg,

    1. Department of Pathology and Laboratory Medicine, University of California, Los Angeles (UCLA), California, USA
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  • Kenneth Kim,

    1. Department of Pathology and Laboratory Medicine, University of California, Los Angeles (UCLA), California, USA
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  • Sam Klein,

    1. Department of Pathology and Laboratory Medicine, University of California, Los Angeles (UCLA), California, USA
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  • Yuhua Zhu,

    1. Department of Pathology and Laboratory Medicine, University of California, Los Angeles (UCLA), California, USA
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  • Chintan Parekh,

    1. Department of Pathology and Laboratory Medicine, University of California, Los Angeles (UCLA), California, USA
    2. Department of Pediatrics, David Geffen School of Medicine (DGSOM), University of California, Los Angeles (UCLA), California, USA
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  • Arumugam Balamurugan,

    1. Department of Medicine, Immunology and Molecular Genetics, Los Angeles (UCLA), CA, USA
    2. Department of Microbiology, Immunology and Molecular Genetics, Los Angeles (UCLA), CA, USA
    3. UCLA AIDS Institute, Los Angeles (UCLA), CA, USA
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  • Otto O. Yang,

    1. Department of Medicine, Immunology and Molecular Genetics, Los Angeles (UCLA), CA, USA
    2. Department of Microbiology, Immunology and Molecular Genetics, Los Angeles (UCLA), CA, USA
    3. UCLA AIDS Institute, Los Angeles (UCLA), CA, USA
    4. AIDS Healthcare Foundation, Los Angeles, CA, USA
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  • Gay M. Crooks

    Corresponding author
    1. Department of Pathology and Laboratory Medicine, University of California, Los Angeles (UCLA), California, USA
    2. Department of Pediatrics, David Geffen School of Medicine (DGSOM), University of California, Los Angeles (UCLA), California, USA
    3. Broad Stem Cell Research Center, Los Angeles (UCLA), CA, USA
    4. Jonsson Comprehensive Cancer Center, University of California, Los Angeles (UCLA), CA, USA
    • Correspondence: Gay M. Crooks, M.B.,B.S. Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California Los Angeles, TLSB, 610 Charles E. Young Drive, East Los Angeles, California 90095-7239, USA. Telephone: 310-206-0205; Fax: 310-206-0356; e-mail: gcrooks@mednet.ucla.edu

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Abstract

A system that allows manipulation of the human thymic microenvironment is needed both to elucidate the extrinsic mechanisms that control human thymopoiesis and to develop potential cell therapies for thymic insufficiency. In this report, we developed an implantable thymic microenvironment composed of two human thymic stroma populations critical for thymopoiesis; thymic epithelial cells (TECs) and thymic mesenchyme (TM). TECs and TM from postnatal human thymi were cultured in specific conditions, allowing cell expansion and manipulation of gene expression, before reaggregation into a functional thymic unit. Human CD34+ hematopoietic stem and progenitor cells (HSPC) differentiated into T cells in the aggregates in vitro and in vivo following inguinal implantation of aggregates in immune deficient mice. Cord blood HSPC previously engrafted into murine bone marrow (BM), migrated to implants, and differentiated into human T cells with a broad T cell receptor repertoire. Furthermore, lentiviral-mediated expression of vascular endothelial growth factor in TM enhanced implant size and function and significantly increased thymocyte production. These results demonstrate an in vivo system for the generation of T cells from human HSPC and represent the first model to allow manipulation of gene expression and cell composition in the microenvironment of the human thymus. Stem Cells 2014;32:2386–2396

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