Reaggregate Thymus Cultures
Andrea White, Eric Jenkinson, Graham Anderson
MRC Center for Immune Regulation, University of Alabama at Birmingham - UAB
Keywords: Immunology, Issue 18, Springer Protocols, Thymus, 2-dGuo, Thymus Organ Cultures, Immune Tolerance, Positive and Negative
Selection, Lymphoid Development,
Date Published: 8/28/2008
Citation: White, A., Jenkinson, E., Anderson, G. Reaggregate Thymus Cultures. J. Vis. Exp. (18), e905, DOI : 10.3791/905 (2008).
In the thymus, immature CD4+8+ thymocytes expressing randomly rearranged T-cell receptor α- and b-chain genes undergo positive and negative
selection events based on their ability to recognize self-peptide/major histocompatibility complex (MHC) molecules expressed by thymic stromal
cells. In vivo analysis of the role of thymic stromal cells during intrathymic selection is made difficult by the cellular complexity of the thymic
microenvironment in the steady-state adult thymus, and by the lack of appropriate targeting strategies to manipulate gene expression in particular
thymic stromal compartments. We have shown that the thymic microenvironment can be readily manipulated in vitro through the use of reaggregate
thymus organ cultures, which allow the preparation of three-dimensional thymus lobes from defined stromal and lymphoid cells. Although other in
vitro systems support some aspects of T-cell development, reaggregate thymus organ culture remains the only in vitro system able to support
efficient MHC class I and II-mediated thymocyte selection events, and so can be used as an effective tool to study the cellular and molecular
regulation of positive and negative selection in the thymus.
The video component of this article can be found at http://www.jove.com/details.php?id=905
For more information on preparing reggregate thymus cultures please visit Springer Protocols.
August 2008 | 18 | e905 | Page 1 of 1
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