Quantifying Human Monocyte Chemotaxis <em>In Vitro</em> and Murine Lymphocyte Trafficking <em>In Vivo</em>

Eliza Prangley; Terrence Kumar; Manish P. Ponda

doi:10.3791/56218

Quantificar o humano Monocyte Chemotaxis In Vitro e linfócitos murino tráfico Vivo

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Quantifying Human Monocyte Chemotaxis In Vitro and Murine Lymphocyte Trafficking In Vivo

Quantificar o humano Monocyte Chemotaxis In Vitro e linfócitos murino tráfico Vivo

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08:38 min

October 30, 2017

DOI: 10.3791/56218-v

Eliza Prangley¹, Terrence Kumar¹, Manish P. Ponda¹

¹Laboratory of Biochemical Genetics and Metabolism,The Rockefeller University

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Overview

This article presents protocols for the quantitative assessment of lymphocyte chemotaxis and migration, crucial for immunology research. It details an in vitro method for real-time evaluation of cell migration and an in vivo technique for tracking native cells to the spleen.

Key Study Components

Area of Science

Immunology
Cell Migration
Chemotaxis

Background

Understanding lymphocyte migration is vital for immunological studies.
Real-time assessment allows for better insights into immune responses.
In vivo tracking provides information on cell behavior within living organisms.
Serum factors can influence immune cell chemotaxis.

Purpose of Study

To quantify monocyte chemotaxis in vitro.
To track lymphocyte migration in vivo.
To explore the effects of serum factors on immune cell behavior.

Methods Used

In vitro labeling of THP-1 cells with fluorescent dye.
Real-time multiplex measurements of cell migration.
In vivo tracking of multiple cell types in recipient animals.
Use of a 35 millimeter cell culture dish for cell preparation.

Main Results

Demonstrated the effectiveness of the in vitro method for real-time analysis.
Showed how specific cell types migrate within a living organism.
Provided quantitative data on the migration of multiple cell types.
Highlighted the influence of serum factors on chemotaxis.

Conclusions

The protocols enhance understanding of lymphocyte behavior.
Real-time assessment tools are valuable for immunology research.
In vivo techniques offer insights into cell migration dynamics.

Frequently Asked Questions

What is the significance of lymphocyte migration?

Lymphocyte migration is crucial for immune responses and understanding disease mechanisms.

How does the in vitro method work?

The in vitro method allows for real-time, quantitative assessment of cell migration using labeled cells.

What are the advantages of in vivo tracking?

In vivo tracking provides insights into how cells behave within the complex environment of a living organism.

What role do serum factors play in chemotaxis?

Serum factors can significantly influence the migration patterns of immune cells.

Can multiple cell types be tracked simultaneously?

Yes, the in vivo method allows for the tracking of multiple cell types within the same animal.

What is the purpose of labeling THP-1 cells?

Labeling THP-1 cells with a fluorescent dye enables visualization and tracking during migration assays.

Protocolos para avaliação quantitativa da quimiotaxia de linfócitos e migração são ferramentas importantes para pesquisa de Imunologia. Aqui, um protocolo em vitro é descrito que permite em tempo real, multiplexados avaliação da migração celular, bem como um complementares na vivo técnica permitindo rastreamento de células nativas para o baço.

Os objetivos gerais desses ensaios são quantificar a quimiotaxia de monócitos in vitro em tempo real e rastrear a migração de linfócitos in vivo. O método in vitro pode ajudar a responder a questões-chave no campo da imunologia, como quais são os efeitos dos fatores séricos na quimiotaxia das células imunes. A principal vantagem da técnica in vitro é que ela permite medições quantitativas e multiplex em tempo real.

O método in vivo pode ajudar a responder a questões-chave no campo da imunologia sobre como tipos específicos de células migram dentro de um animal receptor. A técnica in vivo é particularmente útil para a avaliação quantitativa da migração de vários tipos de células dentro do mesmo animal. Antes de iniciar o experimento, rotule as células THP-1 com um corante fluorescente em uma placa de cultura de células de 35 milímetros em meio completo a 37 graus Celsius por 30 minutos.

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