Preparation of 2-dGuo-Treated Thymus Organ Cultures

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@article{182d3ea039ad4bd1abd7c871962b8953,
title = "Preparation of 2-dGuo-Treated Thymus Organ Cultures",
abstract = "In the thymus, immature CD4+8+ thymocytes expressing randomly rearranged T-cell receptor alpha- 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.",
author = "William Jenkinson and Eric Jenkinson and Graham Anderson",
year = "2008",
month = aug,
day = "28",
doi = "10.3791/906",
language = "English",
journal = "Journal of Visualized Experiments ",
issn = "1940-087X",
publisher = "Journal of Visualized Experiments",
number = "18",

}

RIS

TY - JOUR

T1 - Preparation of 2-dGuo-Treated Thymus Organ Cultures

AU - Jenkinson, William

AU - Jenkinson, Eric

AU - Anderson, Graham

PY - 2008/8/28

Y1 - 2008/8/28

N2 - In the thymus, immature CD4+8+ thymocytes expressing randomly rearranged T-cell receptor alpha- 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.

AB - In the thymus, immature CD4+8+ thymocytes expressing randomly rearranged T-cell receptor alpha- 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.

U2 - 10.3791/906

DO - 10.3791/906

M3 - Article

C2 - 19066498

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 18

ER -