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JoVE Journal
Immunology and Infection
Quantifizierung der menschliche Monocyte Chemotaxis In Vitro und murinen Lymphozyten Men...
Quantifizierung der menschliche Monocyte Chemotaxis In Vitro und murinen Lymphozyten Men...
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

Quantifizierung der menschliche Monocyte Chemotaxis In Vitro und murinen Lymphozyten Menschenhandel In Vivo

Full Text
7,805 Views
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.

Protokolle für die quantitative Bewertung der Lymphozyten Chemotaxis und Migration sind wichtige Werkzeuge für die Immunologie Forschung. Hier wird eine in-vitro- Protokoll beschrieben, dass Genehmigungen in Echtzeit, Multiplex, Bewertung von Zellwanderung sowie eine ergänzende in Vivo Technik ermöglicht Tracking von native Zellen Milz.

Das übergeordnete Ziel dieser Assays ist die Quantifizierung der Monozyten-Chemotaxis in vitro in Echtzeit und die Verfolgung der Lymphozytenmigration in vivo. Die In-vitro-Methode kann helfen, zentrale Fragen im Bereich der Immunologie zu beantworten, z. B. welche Auswirkungen Serumfaktoren auf die Chemotaxis von Immunzellen haben. Der Hauptvorteil der In-vitro-Technik besteht darin, dass sie quantitative und Multiplex-Messungen in Echtzeit ermöglicht.

Die in vivo-Methode kann helfen, zentrale Fragen im Bereich der Immunologie zu beantworten, wie bestimmte Zelltypen innerhalb eines Empfängertieres wandern. Die In-vivo-Technik ist besonders nützlich für die quantitative Beurteilung der Migration mehrerer Zelltypen innerhalb desselben Tieres. Vor Beginn des Experiments werden THP-1-Zellen in einer 35-Millimeter-Zellkulturschale in vollständigem Medium bei 37 Grad Celsius 30 Minuten lang mit einem Fluoreszenzfarbstoff markiert

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