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Related Experiment Videos

Culturing at atmospheric oxygen levels impacts lymphocyte function.

Kondala R Atkuri1, Leonard A Herzenberg, Leonore A Herzenberg

  • 1Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305-5318, USA. atkuri@stanford.edu

Proceedings of the National Academy of Sciences of the United States of America
|March 2, 2005
PubMed
Summary

Culturing immune cells at atmospheric oxygen levels significantly increases T cell proliferation compared to physiologic levels, but only for specific stimuli like CD3/CD28 crosslinking. This finding impacts how researchers study immune responses in vitro.

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Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • In vitro cell culture conditions can significantly impact experimental outcomes.
  • Physiologic oxygen levels (5-10%) are increasingly recognized as crucial for accurately modeling in vivo cellular functions.
  • Atmospheric oxygen (20%) is a common but potentially distorting condition for immune cell culturing.

Purpose of the Study:

  • To investigate the effect of different oxygen concentrations on peripheral blood mononuclear cell (PBMC) responses.
  • To compare lymphocyte proliferation under atmospheric versus physiologic oxygen levels.
  • To determine if oxygen levels influence T cell activation mimicking antigen presentation.

Main Methods:

  • Peripheral blood mononuclear cells were cultured at 5%, 10%, and 20% oxygen for 5 days.

Related Experiment Videos

  • Lymphocyte proliferation was stimulated using Concanavalin A (Con A), anti-CD3/CD28 antibodies, and phytohemagglutinin (PHA).
  • Cell viability was assessed in unstimulated cultures.

    • Main Results:

    • Lymphocyte proliferation increased with rising oxygen levels when stimulated by Con A and anti-CD3/CD28 antibodies.
    • Oxygen levels did not affect proliferation stimulated by phytohemagglutinin.
    • Cell viability remained unaffected by oxygen levels in unstimulated cultures.

    Conclusions:

    • The impact of oxygen levels on lymphocyte proliferation is stimulus-dependent.
    • Culturing cells at atmospheric oxygen significantly enhances proliferation in response to CD3/CD28 crosslinking, a common mimic of T cell receptor signaling.
    • These findings highlight the importance of using physiologic oxygen levels for accurate in vitro immune response studies.