Neutrophil Isolation and Analysis to Determine their Role in Lymphoma Cell Sensitivity to Therapeutic Agents

Taghreed Hirz; Charles Dumontet

doi:10.3791/53846

Aislamiento de neutrófilos y análisis para determinar su papel en el linfoma de células sensibles...

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Immunology and Infection

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JoVE Journal Immunology and Infection

Neutrophil Isolation and Analysis to Determine their Role in Lymphoma Cell Sensitivity to Therapeutic Agents

Aislamiento de neutrófilos y análisis para determinar su papel en el linfoma de células sensibles a los agentes terapéuticos

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14:04 min

March 25, 2016

DOI: 10.3791/53846-v

Taghreed Hirz¹, Charles Dumontet¹

¹Immunity, Microenvironment, Virus,Cancer Research Center in Lyon

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Overview

This article describes the isolation of neutrophils from human blood using density gradient separation and the differentiation of HL60 cells along the granulocytic pathway. The study aims to investigate the role of neutrophils in the sensitivity of lymphoma cells to anti-lymphoma agents.

Key Study Components

Area of Science

Immunology
Oncology
Cell Biology

Background

Neutrophils are the most abundant type of white blood cells.
They play a crucial role in the innate immune response.
Understanding their interaction with cancer cells can inform therapeutic strategies.
Density gradient centrifugation allows for the isolation of pure immune cell populations.

Purpose of Study

To isolate highly purified neutrophils from human blood samples.
To explore the role of neutrophils in lymphoma cell sensitivity to cancer therapy.
To utilize both 2D and 3D cell culture models for investigation.

Methods Used

Density gradient centrifugation for neutrophil isolation.
Use of EDTA treated blood from chronic lymphocytic leukemia patients.
Preparation of blood samples with RPMI dilution.
Layering of density gradient solution to separate cell types.

Main Results

Successful isolation of neutrophils without altering their function.
Insights into the interactions between neutrophils and lymphoma cells.
Potential implications for enhancing cancer therapy effectiveness.
Establishment of methods for future research in cancer immunology.

Conclusions

Neutrophils may significantly influence lymphoma cell responses to treatment.
Density gradient separation is an effective method for isolating immune cells.
Further studies are needed to fully understand neutrophil roles in cancer therapy.

Frequently Asked Questions

What are neutrophils?

Neutrophils are the most abundant type of white blood cells and play a key role in the immune response.

How are neutrophils isolated from blood?

Neutrophils can be isolated using density gradient centrifugation, which separates cells based on their density.

What is the significance of studying neutrophils in cancer?

Studying neutrophils can provide insights into their role in tumor responses and potential therapeutic strategies.

What types of cell culture models are used in this study?

Both 2D and 3D cell culture models are utilized to investigate neutrophil interactions with lymphoma cells.

What is the main goal of this research?

The main goal is to understand how neutrophils affect the sensitivity of lymphoma cells to anti-lymphoma agents.

What are the advantages of density gradient separation?

It allows for the rapid isolation of pure immune cell populations without altering their function.

What future research directions does this study suggest?

Future research may focus on the detailed mechanisms of neutrophil-mediated tumor responses and their implications for therapy.

Los neutrófilos son el tipo más abundante de glóbulos blancos. Aquí se describe el aislamiento de neutrófilos de la sangre humana mediante el método de separación en gradiente de densidad y la diferenciación de células promielocíticas humanas (HL60) a lo largo de la vía granulocítica; para probar su papel en la sensibilidad de las células de linfoma a los agentes antilinfomas.

El objetivo general de estos procedimientos es aislar rápidamente neutrófilos altamente purificados de muestras de sangre humana mediante centrifugación en gradiente de densidad para investigar su papel en la sensibilidad de las células de linfoma a la terapia contra el cáncer utilizando dos modelos de cultivo de células D y tres D. Estos métodos pueden ayudar a responder preguntas clave sobre las interacciones entre el sistema inmunitario innato y el cáncer, como qué papel desempeñan las respuestas de las células tumorales mediadas por neutrófilos en la terapia contra el cáncer. La principal ventaja de la separación por gradiente de densidad es la capacidad de aislar poblaciones de células inmunitarias puras en un corto período de tiempo sin exponer las células a estímulos que alteren la función.

Para aislar las células leucémicas primarias y los neutrófilos, comience por agregar 15 mililitros de sangre tratada con EDTA de muestras de pacientes con leucemia linfocítica crónica en tubos individuales y estériles de 50 mililitros. Diluye la sangre con 15 mililitros de RPMI. Luego, agregue con cuidado y lentamente 15 mililitros de solución de gradiente de densidad a temperatura ambiente debajo de la sangre, teniendo cuidado de no mezclar las capas.

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