ERRATUM NOTICE
Important: There has been an erratum issued for this article. Read more …
Summary
이 비디오에의 준비 2 dGuo - 처리 증명 thymus 문화를 reaggregate.
Abstract
thymus, 미숙 CD4 8 + 무작위로 재배열 T - 세포 수용체를 표현 thymocytes에서 α - 및 B - 체인 유전자 self-peptide/major의 histocompatibility 복합 (MHC) thymic로 표현 분자를 인식하는 능력에 따라 긍정적이고 부정적인 선택 이벤트를 받다 stromal 세포. intrathymic 선택하는 동안 thymic stromal 세포의 역할의 생체내 분석에 정상 상태 성인 thymus에서 thymic microenvironment의 세포 복잡하여 어렵게 만든, 적절한 타겟팅 전략의 부족에 의해 특정 thymic stromal 구획의 유전자 발현을 조작할 수 있습니다. 우리는 thymic microenvironment가 쉽게 정의 stromal과 림프 세포의 입체 thymus 엽 (叶)의 준비를 허용 thymus 장기 문화를 reaggregate의 사용을 통해 체외로 조작할 수있는 것으로 나타났습니다. 체외 시스템의 다른 T 세포 개발의 일부 측면을 지원하지만, thymus 장기 문화 I 및 II - 중재 thymocyte 선택 이벤트, 그래서이 연구는 효과적인 도구로 사용할 수있는 유일한 체외 시스템의 효율적인 MHC 클래스를 지원할 수 남아 reaggregate thymus에서 긍정적이고 부정적인 선택의 세포 및 분자 규제.
Protocol
thymus 문화를 reggregate 준비에 대한 자세한 내용은 방문하시기 바랍니다 스프링거 프로토콜을 .
Tags
면역학 제 18 스프링거 프로토콜 Thymus 2 dGuo Thymus의 오르간 문화 면역 공차 긍정적이고 부정적인 선택 림프 개발Erratum
Formal Correction: Erratum: Reaggregate Thymus Cultures
Posted by JoVE Editors on 04/01/2012.
Citeable Link.
A correction was made to: Reaggregate Thymus Cultures. A revised abstract was republished due to a publisher error. The abstract was corrected to:
Stromal cells within lymphoid tissues are organized into three-dimensional structures that provide a scaffold that is thought to control the migration and development of haemopoeitic cells. Importantly, the maintenance of this three-dimensional organization appears to be critical for normal stromal cell function, with two-dimensional monolayer cultures often being shown to be capable of supporting only individual fragments of lymphoid tissue function. In the thymus, complex networks of cortical and medullary epithelial cells act as a framework that controls the recruitment, proliferation, differentiation and survival of lymphoid progenitors as they undergo the multi-stage process of intrathymic T-cell development. Understanding the functional role of individual stromal compartments in the thymus is essential in determining how the thymus imposes self/non-self discrimination. Here we describe a technique in which we exploit the plasticity of fetal tissues to re-associate into intact three-dimensional structures in vitro, following their enzymatic disaggregation. The dissociation of fetal thymus lobes into heterogeneous cellular mixtures, followed by their separation into individual cellular components, is then combined with the in vitro re-association of these desired cell types into three-dimensional reaggregate structures at defined ratios, thereby providing an opportunity to investigate particular aspects of T-cell development under defined cellular conditions. (This article is based on work first reported Methods in Molecular Biology 2007, Vol. 380 pages 185-196).
from
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.
