Name: reaggregate thymus cultures
Uploaded: 2008-08-28T13:00:00
Duration: 5 min 47 s
Description: Watch this Scientific Journal Video about Reaggregate Thymus Cultures at JoVE.com

तैयारी thymus संस्कृतियों reggregate के बारे में अधिक जानकारी के लिए कृपया पर <a href="http://www.springerprotocols.com/">जाएँ स्प्रिंगर</a> प्रोटोकॉल .

The authors have nothing to disclose.

A correction was made to: <a href="http://www.jove.com/video/905/">Reaggregate Thymus Cultures.</a> 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 &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/905/">Reaggregate Thymus Cultures</a>. A revised abstract was republished due to a publisher error.

Erratum: Reaggregate Thymus Cultures

reaggregate thymus cultures

Thymus, अपरिपक्व 8 CD4 + बेतरतीब ढंग से पुनर्व्यवस्थित टी सेल रिसेप्टर व्यक्त thymocytes में α और ख श्रृंखला जीन सकारात्मक और नकारात्मक चयन उनके self-peptide/major उतक अनुरूपता (MHC) जटिल thymic द्वारा व्यक्त अणुओं की पहचान करने की क्षमता के आधार पर की घटनाओं से गुजरना stromal कोशिकाओं. Intrathymic चयन के दौरान thymic stromal कोशिकाओं की भूमिका के vivo विश्लेषण में स्थिर राज्य वयस्क thymus में thymic microenvironment के सेलुलर जटिलता से मुश्किल बना है, और उपयुक्त लक्ष्यीकरण रणनीतियों की कमी से विशेष thymic stromal डिब्बों में जीन की अभिव्यक्ति में हेरफेर करने के लिए. हम पता चला है कि thymic microenvironment reaggregate thymus अंग संस्कृतियों, जो परिभाषित stromal और lymphoid कोशिकाओं से तीन आयामी thymus lobes के तैयारी की अनुमति के उपयोग के माध्यम से आसानी से किया जा सकता है इन विट्रो में हेरफेर. हालांकि इन विट्रो प्रणालियों में अन्य टी सेल विकास के कुछ पहलुओं का समर्थन reaggregate, thymus अंग संस्कृति इन विट्रो प्रणाली में ही रहता है कुशल MHC वर्ग का समर्थन करने में सक्षम मैं और द्वितीय की मध्यस्थता thymocyte चयन घटनाओं, और इसलिए अध्ययन करने के लिए एक प्रभावी उपकरण के रूप में इस्तेमाल किया जा सकता है thymus में सकारात्मक और नकारात्मक चयन के सेलुलर और आणविक विनियमन.

Watch this Scientific Journal Video about Reaggregate Thymus Cultures at JoVE.com

Reaggregate Thymus Cultures

इस वीडियो की तैयारी 2 dGuo इलाज reaggregate thymus संस्कृतियों का प्रदर्शन.

Thymus संस्कृति Reaggregate

Stromal cells within lymphoid tissues are organized into three-dimensional structures that provide a scaffold that is thought to control the migration and ...

Research

JoVE Journal

Biology

ES Cell-derived Neuroepithelial Cell Cultures

ES सेल व्युत्पन्न Neuroepithelial सेल संस्कृतियों

ES cells have the potential to differentiate into cells from all germ layers, which makes them an attractive tool for the development of new therapies. In ...

Preparation of Neuronal Cultures from Midgastrula Stage Drosophila Embryos

Midgastrula स्टेज ड्रोसोफिला भ्रूण से Neuronal संस्कृति की तैयारी

This video illustrates the procedure for making primary neuronal cultures from midgastrula stage Drosophila embryos. The methods for collecting embryos ...

Preparation of Dissociated Mouse Cortical Neuron Cultures

Dissociated माउस cortical न्यूरॉन संस्कृति की तैयारी

This video will guide you through the process for generating cortical neuronal cultures from late embryo and early postnatal mouse brain. These cultures ...

In situ Imaging of the Mouse Thymus Using 2-Photon Microscopy

माउस 2-photon माइक्रोस्कोपी का उपयोग thymus के स्वस्थानी इमेजिंग में

Two-photon Microscopy (TPM) enables us to image deep into the thymus and document the events that are important for thymocyte development.  To follow the ...

Mouse Epidermal Neural Crest Stem Cell (EPI-NCSC) Cultures

Mouse Epidermal Neural Crest Stem Cell EPI-NCSC Cultures

माउस Epidermal तंत्रिका शिखा स्टेम सेल (महामारी NCSC) संस्कृति

EPI-NCSC are remnants of the embryonic neural crest in an adult location, the bulge of hair follicles. They are multipotent stem cells that have the ...

Primary Dissociated Midbrain Dopamine Cell Cultures from Rodent Neonates

प्राथमिक dissociated कृंतक नवजात शिशुओं से midbrain dopamine सेल संस्कृतियों

The ability to create primary cell cultures of dopamine neurons allows for the study of the presynaptic characteristics of dopamine neurons in isolation ...

Preparation of 2-dGuo-Treated Thymus Organ Cultures

2 - dGuo इलाज thymus अंग संस्कृतियों की तैयारी

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

Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

की तैयारी Aplysia संवेदी मोटर neuronal सेल संस्कृतियों

The nervous system of the marine mollusk Aplysia californica is relatively simple, consisting of approximately 20,000 neurons.  The neurons are large (up ...

Establishing Primary Adult Fibroblast Cultures From Rodents

कृन्तकों से प्राथमिक वयस्क fibroblast संस्कृतियों की स्थापना

The importance of using primary cells, rather than cancer cell lines, for biological studies is becoming widely recognized.  Primary cells are preferred ...

Primary Cell Cultures from Drosophila Gastrula Embryos

Primary Cell Cultures from Drosophila Gastrula Embryos

प्राथमिक सेल संस्कृति से ड्रोसोफिला गेसट्रुला भ्रूण

Here we describe a method for preparing and culturing primary cells dissociated from Drosophila gastrula embryos.   In brief, a large amount of staged ...