ERRATUM NOTICE
Important: There has been an erratum issued for this article. Read more …
Summary
在这个视频的准备2 dGuo治疗reaggregate胸腺文化是证明。
Abstract
在胸腺，不成熟的CD4 + 8 +胸腺细胞表达随机重排的T细胞受体的α-和B链的基因进行积极和消极的选择根据自己的能力，认识到self-peptide/major胸腺表达的组织相容性复合体（MHC）分子的事件基质细胞。在体内的胸腺基质细胞在胸腺内选择的作用分析是难以在胸腺微环境的稳态成人胸腺细胞的复杂性，以及缺乏适当的定位策略来操纵基因的表达，尤其是胸腺基质车厢。我们已经证明胸腺微环境，可以在体外很容易操纵，通过使用reaggregate胸腺器官培养，这让准备从定义的基质和淋巴样细胞的三维胸腺叶。虽然在体外系统支持T细胞发育的某些方面，reaggregate胸腺器官培养仍然是唯一在体外系统能够支持高效的MHCⅠ类，我和II介导的胸腺细胞的选择事件，等等，可作为一种有效的工具来研究使用的正面和负面的选择在胸腺细胞和分子调控。
Protocol
对于准备reggregate胸腺文化的更多信息， 请访问斯普林格协议。
Tags
免疫学，第18期，2 dGuo，斯普林格协议，胸腺，胸腺器官培养，免疫耐受，积极和消极选择，淋巴发展Erratum
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.
