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JoVE Journal
Immunology and Infection
ひと単球走化性の in VitroおよびIn Vivoマウス リンパ球の定量化
ひと単球走化性の in VitroおよびIn Vivoマウス リンパ球の定量化
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

ひと単球走化性の in VitroおよびIn Vivoマウス リンパ球の定量化

Full Text
7,810 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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Please note that some of the translations on this page are AI generated. Click here for the English version.

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.

リンパ球走化性と移行の定量的評価のためのプロトコルは、免疫学の研究のための重要なツールです。ここでは、リアルタイムで、多重セル移行の評価だけでなく、生体内で相補的な技術有効にする追跡ネイティブ細胞の脾臓に、体外のプロトコルを説明します。

これらのアッセイの全体的な目標は、単球の走化性をin vitroでリアルタイムに定量化し、in vivoでリンパ球の遊走を追跡することです。in vitro法は、免疫細胞の走化性に対する血清因子の影響など、免疫学の分野における重要な質問に答えるのに役立ちます。in vitro技術の主な利点は、リアルタイムでの定量的および多重測定が可能なことです。

in vivo法は、特定の細胞タイプがレシピエント動物内でどのように移動するかという免疫学の分野における重要な質問に答えるのに役立ちます。in vivo技術は、同じ動物内の複数の細胞タイプの移動の定量的評価に特に有用です。実験を開始する前に、35 mm細胞培養皿中の蛍光色素でTHP-1細胞を完全培地中で摂氏37度、30分間標識します。

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