Name: preparation of 2-dguo-treated thymus organ cultures
Uploaded: 2008-08-28T19:00:00
Duration: 7 min 50 s
Description: Watch this Scientific Journal Video about Preparation of 2-dGuo-Treated Thymus Organ Cultures at JoVE.com

يرجى زيارة <a href="http://www.springerprotocols.com/">البروتوكولات سبرينغر</a> للحصول على مزيد من المعلومات حول إعداد الجهاز السابقين فيفو الثقافات التوتة.

The authors have nothing to disclose.

A correction was made to: <a href="http://www.jove.com/video/906/">Preparation of 2-dGuo-Treated Thymus Organ Cultures</a>. A revised abstract was republished due to a publisher error. The abstract was corrected to:
In the thymus, interactions between developing T-cell precursors and stromal cells that include cortical and medullary epithelial cells are known to play a key role in the development of a functionally competent T-cell pool. However, the complexity of T-cell development in the thymus in vivo can limit analysis of individual cellular components and particular stages of development. In vitro culture systems provide a readily accessible means to study multiple complex cellular processes. Thymus organ culture systems represent a widely used approach to study intrathymic development of T-cells under defined conditions in vitro. Here we describe a system in which mouse embryonic thymus lobes can be depleted of endogenous haemopoeitic elements by prior organ culture in 2-deoxyguanosine, a compound that is selectively toxic to haemopoeitic cells. As well as providing a readily accessible source of thymic stromal cells to investigate the role of thymic microenvironments in the development and selection of T-cells, this technique also underpins further experimental approaches that include the reconstitution of alymphoid thymus lobes in vitro with defined haemopoietic elements, the transplantation of alymphoid thymuses into recipient mice, and the formation of reaggregate thymus organ cultures. (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 &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.

A correction was made to: <a href="http://www.jove.com/video/906/">Preparation of 2-dGuo-Treated Thymus Organ Cultures</a>. A revised abstract was republished due to a publisher error.

Erratum: Preparation of 2-dGuo-Treated Thymus Organ Cultures

preparation of 2-dguo-treated thymus organ cultures

في CD4 ، الغدة الصعترية غير ناضجة +8 + thymocytes معربا عن ترتيبها عشوائيا مستقبلات الخلايا T - α وب سلسلة الجينات يخضع اختيار الأحداث الإيجابية والسلبية على أساس قدرتهم على الاعتراف self-peptide/major معقدة التوافق النسيجي (MHC) الجزيئات التي أعربت عنها التوتة انسجة الخلايا. في تحليل لدور الجسم الحي من خلايا انسجة الغدة الصعترية خلال اختيار intrathymic يرصد صعبة بسبب تعقيد الخلوية من المكروية التوتة التوتة الكبار في حالة استقرار ، وبسبب عدم وجود استراتيجيات مناسبة لمعالجة استهداف الجينات في مقصورات خاصة انسجة الغدة الصعترية. لقد أظهرنا أنه لا يمكن التلاعب بها بسهولة المكروية التوتة في المختبر من خلال استخدام جهاز التوتة reaggregate الثقافات ، والتي تتيح إعداد ثلاثي الأبعاد فصوص من الغدة الصعترية تعريف الخلايا اللحمية واللمفاوية. على الرغم من أن الأنظمة الأخرى في المختبر دعم بعض جوانب تي خلية التنمية ، reaggregate التوتة الثقافة الجهاز يبقى فقط في نظام المختبر قادرا على دعم الطبقة MHC فعالة يمكن استخدامها الأول والثاني بوساطة الأحداث اختيار خلية توتية ، وذلك باعتبارها أداة فعالة لدراسة تنظيم الخلوية والجزيئية لاختيار الإيجابية والسلبية في الغدة الصعترية.

Watch this Scientific Journal Video about Preparation of 2-dGuo-Treated Thymus Organ Cultures at JoVE.com

Preparation of 2-dGuo-Treated Thymus Organ Cultures

هذا الفيديو يوضح تشريح وازالة الغدة الصعترية الجنين وكذلك التحضير للثقافات فيفو السابقين من 2 dGuo المعالجة الغدة الصعترية.

إعداد 2 - dGuo المعاملة الثقافات الجهاز الغدة الصعترية

In the thymus, interactions between developing T-cell precursors and stromal cells that include cortical and medullary epithelial cells are known to play ...

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