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Human plasma-like medium improves T lymphocyte activation

View ORCID ProfileMichael A. Leney-Greene, Arun K. Boddapati, Helen C. Su, Jason Cantor, Michael J. Lenardo
doi: https://doi.org/10.1101/740845
Michael A. Leney-Greene
1Molecular Development of the Immune System Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892
2Immunology Graduate Group, Biomedical Graduate Studies, University of Pennsylvania, Philadelphia, PA 19104
7Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706
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  • ORCID record for Michael A. Leney-Greene
Arun K. Boddapati
3NIAID Collaborative Bioinformatics Resource (NCBR), National Institute of Allergy and infectious Diseases, National Institutes of Health, Bethesda, MD, United States
4Advanced Biomedical Computational Science, Frederick National Laboratory for Cancer Research, Frederick, MD, United States
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Helen C. Su
2Immunology Graduate Group, Biomedical Graduate Studies, University of Pennsylvania, Philadelphia, PA 19104
5Human Immunological Diseases Section, Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892
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Jason Cantor
6Morgridge Institute for Research, 330 North Orchard Street, Madison, WI 53715
7Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706
8Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53706
9Carbone Cancer Center, University of Wisconsin-Madison, 600 Highland Avenue, Madison, WI 53705
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Michael J. Lenardo
1Molecular Development of the Immune System Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892
2Immunology Graduate Group, Biomedical Graduate Studies, University of Pennsylvania, Philadelphia, PA 19104
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  • For correspondence: michael.lenardo@nih.gov
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SUMMARY

T lymphocytes are critical for effective immunity and the ability to study their behavior in synthetic media in vitro facilitates major discoveries in their development, function, and fate. However, the composition of human plasma differs from synthetic media and we hypothesized that these differences could have important effects on cell physiology. We therefore compared T lymphocyte activation in human plasma-like medium (HPLM) to RPMI supplemented with dialyzed FBS (RPMIdFBS) and found that it entrained markedly different transcriptional responses. We also found that the concentration of calcium in RPMIdFBS is six-fold lower than HPLM causing altered T cell activation which could be reversed by calcium addition. Thus, investigators should be cognizant of differences between commonly used media formulations and HPLM which is based on the in vivo plasma environment as these could profoundly affect their experimental results. Physiologic media may be a valuable new way to study immune cells in culture.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. This article is a US Government work. It is not subject to copyright under 17 USC 105 and is also made available for use under a CC0 license.
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Posted August 21, 2019.
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Human plasma-like medium improves T lymphocyte activation
Michael A. Leney-Greene, Arun K. Boddapati, Helen C. Su, Jason Cantor, Michael J. Lenardo
bioRxiv 740845; doi: https://doi.org/10.1101/740845
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Human plasma-like medium improves T lymphocyte activation
Michael A. Leney-Greene, Arun K. Boddapati, Helen C. Su, Jason Cantor, Michael J. Lenardo
bioRxiv 740845; doi: https://doi.org/10.1101/740845

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