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

Quantifier la chimiotaxie Monocyte humain In Vitro et lymphocytes murins traite In V...

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Immunology and Infection

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JoVE Journal Immunology and Infection

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

Quantifier la chimiotaxie Monocyte humain In Vitro et lymphocytes murins traite In 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.

Protocoles pour une évaluation quantitative de la chimiotaxie des lymphocytes et des migrations sont des outils importants pour la recherche de l’immunologie. Un protocole in vitro est décrit ici, que les permis en temps réel, multiplexés évaluation de la migration cellulaire, ainsi qu’un complémentaire en vivo technique propice suivi de cellules natives de rate.

Les objectifs globaux de ces tests sont de quantifier la chimiotaxie des monocytes in vitro en temps réel et de suivre la migration des lymphocytes in vivo. La méthode in vitro peut aider à répondre à des questions clés dans le domaine de l’immunologie, telles que les effets des facteurs sériques sur la chimiotaxie des cellules immunitaires. Le principal avantage de la technique in vitro est qu’elle permet des mesures quantitatives et multiplex en temps réel.

La méthode in vivo peut aider à répondre à des questions clés dans le domaine de l’immunologie sur la façon dont des types de cellules spécifiques migrent au sein d’un animal receveur. La technique in vivo est particulièrement utile pour l’évaluation quantitative de la migration de plusieurs types de cellules au sein d’un même animal. Avant de commencer l’expérience, étiquetez les cellules THP-1 avec un colorant fluorescent dans une boîte de culture cellulaire de 35 millimètres dans un milieu complet à 37 degrés Celsius pendant 30 minutes.

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