Un abonnement à JoVE est nécessaire pour voir ce contenu.  Connectez-vous ou commencez votre essai gratuit.
Determining the Role of Neutrophil-Like Cells in Lymphoma Cell Sensitivity to a Test Drug

Determining the Role of Neutrophil-Like Cells in Lymphoma Cell Sensitivity to a Test Drug

Transcription

Take a multi-well plate containing 3D spheroid monocultures of lymphoma cells, cancerous white blood cells, and co-cultures of lymphoma and neutrophil-like cells in a basement membrane matrix.

Add a test drug that binds to intracellular tubulin and disrupts microtubule polymerization, resulting in cellular apoptosis.

In the co-culture, direct lymphoma cell contact activates the neutrophil-like cells, inducing cytokine secretion.

These cytokines interact with lymphoma cell receptors, triggering downstream signaling cascades and upregulating anti-apoptotic protein expression, promoting cell survival.

Discard the media. Add a dissociation buffer to disrupt cellular adhesion.

Scrape the spheroids and incubate them under agitation. Transfer the suspension to tubes and continue agitation to form single-cell suspensions.

Introduce a fluorophore-conjugated antibody cocktail that labels lymphoma and neutrophil-like cells.

Add fluorophore-tagged annexin V and propidium iodide to label apoptotic and dead cells.

Using flow cytometry, detect increased lymphoma cell survival in the co-culture, indicating neutrophil-mediated protection against the drug.

Vidéos Connexes

Read Article