Name: generation of bone marrow derived murine dendritic cells for use in 2-photon imaging
Uploaded: 2008-07-09T16:32:00
Description: Watch this Scientific Journal Video about Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging at JoVE.com

National Institutes of Health Kirchstein Fellowship predoctorale gemeenschap AI-64128 (MPM), GM-41514 (MDC), GM-48.071 (IP)

1) Verwijder beide dijbeen botten van een muis <ol><li> Gebruik dissectie schaar weg te snijden spieren en het dijbeen bot bloot boven en onder de gewrichten (knie en heup). Pak het midden van het dijbeen met de dissectie een pincet en snijd boven en onder de gewrichten om zoveel mogelijk van de epifyse als mogelijk intact laten. </li><li> Schoon uit zo veel spieren als mogelijk met behulp van kleine dissectie schaar. Overdracht het dijbeen in een schaal van RPMI. </li><li> Alle procedures moeten worden uitgevoerd in d...

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Dendritische cellen zijn de belangrijkste bemiddelaars van de adaptieve immuunrespons en de meest efficiënte antigeen presenterende cel gekenmerkt to-date. Methoden voor mens en muis dendritische cel-cultuur in de literatuur verschillen in de aard van de cytokines gebruikt om de ontwikkeling van de verschillende soorten dendritische cel beïnvloeden. Met name zijn GM-CSF, flt3L, IL4, IL13, TNF-alfa en IFN-gamma gebruikt in verschillende combinaties te produceren volwassen, onvolwassen, ontstekings-en steady-state, zoals muizen beenmerg afg...

generation of bone marrow derived murine dendritic cells for use in 2-photon imaging

Verschillende methoden voor de bereiding van muizen dendritische cellen kan worden gevonden in de literatuur. Hier presenteren we een methode die meer dan 85% CD11c hoge dendritische cellen produceert in cultuur die de thuishaven van de drainerende lymfeklier na subcutane injectie en presenteren antigeen aan antigeen specifieke T-cellen (zie video). Daarnaast maken we gebruik van Essen Instruments Incucyte naar dendritische cel rijping, waar, op dag 10, de morfologie van de gekweekte cellen is typerend voor een rijpe dendritische cel en &lt;85% van de cellen zijn CD11chigh track. De studie van antigeen presentatie in perifere lymfeklieren door twee-foton beeldvorming bleek dat er drie verschillende fasen van dendritische cel en T-cel interactie 1, 2. Fase I bestaat uit korte seriële contacten tussen zeer beweeglijke antigeen specifieke T cellen en antigen dragen dendritische cellen 1, 2. Fase twee wordt gekenmerkt door langdurige contacten tussen antigeen-specifieke T-cel en antigeen met dendritische cellen 1, 2. Tot slot wordt fase III kenmerkt zich door T-cellen losmaken van dendritische cellen, het herwinnen van de beweeglijkheid en het begin tot het 1, 2 delen. Dit is een voorbeeld van het type van antigeen-specifieke interacties die kunnen worden geanalyseerd door twee-foton beeldvorming van antigeen-beladen cellen tracker dye-gelabelde dendritische cellen.

Watch this Scientific Journal Video about Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging at JoVE.com

Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging

Antigeen presentatie in secundaire lymfoïde organen door de dendritische cellen is cruciaal voor de opening van het T-cel gemedieerde adaptieve immuunrespons. Hier laten we zien de cultuur van beenmerg afgeleide dendritische cellen van muizen, activering, en de etikettering voor 2-photon imaging.

Generatie van beenmerg afgeleide Murine dendritische cellen voor gebruik in twee-foton Imaging

Several methods for the preparation of murine dendritic cells can be found in the literature. Here, we present a method that produces greater than 85% ...

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Biology

Dissection and 2-Photon Imaging of Peripheral Lymph Nodes in Mice

Dissection en 2-Photon Imaging van perifere lymfeklieren in Muizen

Two-photon imaging has revealed an elegant choreography of motility and cellular interactions within the lymph node under basal conditions and at the ...

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

In situ beeldvorming van de muis Thymus Met behulp van 2-Photon Microscopie

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 ...

A Method for 2-Photon Imaging of Blood Flow in the Neocortex through a Cranial Window

Een methode voor 2-Photon Imaging van de bloedstroom in de neocortex door middel van een Cranial Window

The ability to image the cerebral vasculature (from large vessels to capillaries) and record blood flow dynamics in the intact brain of living rodents is ...

In Vivo 2-Photon Calcium Imaging in Layer 2/3 of Mice

In vivo 2-Photon Imaging Calcium in Layer 2 / 3 van Muizen

To understand network dynamics of microcircuits in the neocortex, it is essential to simultaneously record the activity of a large number of neurons .  ...

Culture of myeloid dendritic cells from bone marrow precursors

Cultuur van myeloïde dendritische cellen uit beenmerg precursors

Myeloid dendritic cells (DCs) are frequently used to study the interactions between innate and adaptive immune mechanisms and the early response to ...

The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

De vervaardiging van primaire hematopoietische cellen culturen uit Murine Bone Marrow voor Elektroporatie

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser ...

Homing of Hematopoietic Cells to the Bone Marrow

Sturen van hematopoietische cellen naar het beenmerg

Homing is the phenomenon whereby transplanted hematopoietic cells are able to travel to and engraft or establish residence in the bone marrow.  Various ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-dimensionale Tracking van hematopoietische stamcellen en progenitorcellen in volwassen muis calvarial Bone Marrow

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Osteoclast Derivation from Mouse Bone Marrow

Osteoclast Afleiding van Mouse Bone Marrow

Osteoclasts are highly specialized cells that are derived from the monocyte/macrophage lineage of the bone marrow. Their unique ability to resorb both the ...

Expression of Fluorescent Fusion Proteins in Murine Bone Marrow-derived Dendritic Cells and Macrophages

Expressie van fluorescerende Fusion eiwitten in lymfkliertest beenmerg-afgeleide dendritische cellen en macrofagen

Dendritic cells and macrophages are crucial cells that form the first line of defense against pathogens. They also play important roles in the initiation ...