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Construction of Thymus Organoids from Decellularized Thymus Scaffolds

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Book cover Organoids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1576))

Abstract

One of the hallmarks of modern medicine is the development of therapeutics that can modulate immune responses, especially the adaptive arm of immunity, for disease intervention and prevention. While tremendous progress has been made in the past decades, manipulating the thymus, the primary lymphoid organ responsible for the development and education of T lymphocytes, remains a challenge. One of the major obstacles is the difficulty to reproduce its unique extracellular matrix (ECM) microenvironment that is essential for maintaining the function and survival of thymic epithelial cells (TECs), the predominant population of cells in the thymic stroma. Here, we describe the construction of functional thymus organoids from decellularized thymus scaffolds repopulated with isolated TECs. Thymus decellularization was achieved by freeze–thaw cycles to induce intracellular ice crystal formation, followed by detergent-induced cell lysis. Cellular debris was removed with extensive wash. The decellularized thymus scaffolds can largely retain the 3D extracellular matrix (ECM) microenvironment that can support the recolonization of TECs. When transplanted into athymic nude mice, the reconstructed thymus organoids can effectively promote the homing of bone marrow-derived lymphocyte progenitors and support the development of a diverse and functional T cell repertoire. Bioengineering of thymus organoids can be a promising approach to rejuvenate/modulate the function of T-cell mediated adaptive immunity in regenerative medicine.

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Acknowledgements

This work was supported in part by the National Institutes of Health grant R01 AI123392 (YF) and by the generous support of Allegheny Health Network to the Institute of Cellular Therapeutics.

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Authors and Affiliations

  1. Institute of Cellular Therapeutics, Allegheny Health Network, Pittsburgh, PA, USA

    Asako Tajima, Isha Pradhan & Massimo Trucco

  2. Department of Dermatology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    Xuehui Geng

  3. Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA, USA

    Massimo Trucco & Yong Fan Ph.D.

  4. Department of Microbiology and Immunology, Medical College of Drexel University, Philadelphia, PA, USA

    Massimo Trucco & Yong Fan Ph.D.

  5. Institute of Cellular Therapeutics, Allegheny Health Network, Room 1107 South Tower, 320 East North Avenue, Pittsburgh, PA, 15212, USA

    Yong Fan Ph.D.

Authors
  1. Asako Tajima
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  2. Isha Pradhan
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  3. Xuehui Geng
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  4. Massimo Trucco
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  5. Yong Fan Ph.D.
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Corresponding author

Correspondence to Yong Fan Ph.D. .

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Editors and Affiliations

  1. Ottawa Hospital Research Institute, Ottawa, ON, Canada

    Kursad Turksen

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© 2016 Springer Science+Business Media New York

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Tajima, A., Pradhan, I., Geng, X., Trucco, M., Fan, Y. (2016). Construction of Thymus Organoids from Decellularized Thymus Scaffolds. In: Turksen, K. (eds) Organoids. Methods in Molecular Biology, vol 1576. Humana, New York, NY. https://doi.org/10.1007/7651_2016_9

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  • DOI: https://doi.org/10.1007/7651_2016_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-7616-4

  • Online ISBN: 978-1-4939-7617-1

  • eBook Packages: Springer Protocols

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