| Original language | English |
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| Article number | 101204 |
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| Journal | STAR Protocols |
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| Volume | 3 |
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| Issue number | 1 |
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| Early online date | 25 Feb 2022 |
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| DOIs | |
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| E-pub ahead of print | 25 Feb 2022 |
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| Published | 18 Mar 2022 |
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Funding Information:
This research was funded by the Fetal Medicine Foundation, United Kingdom and supported by the National Institute for Health Research (NIHR) Biomedical Research Centre based at Guy's and St Thomas? NHS Foundation Trust and King's College London and/or the NIHR Clinical Research Facility. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care. The graphical abstract was created with Biorender.com. A.E. and N.M. consented patients, collected, processed the blood, and analyzed the data. P.S. processed blood, analyzed data, and provided expert advice. R.P. analyzed the blood and data. S.T.M. wrote and edited the manuscript. D.M. C.S. G.L. K.H.N. provided expert advice. All authors approved the manuscript. The authors declare no competing interests.
Funding Information:
This research was funded by the Fetal Medicine Foundation , United Kingdom and supported by the National Institute for Health Research (NIHR) Biomedical Research Centre based at Guy’s and St Thomas’ NHS Foundation Trust and King’s College London and/or the NIHR Clinical Research Facility. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care. The graphical abstract was created with Biorender.com .
Publisher Copyright:
© 2022 The Authors
To analyze immune cell populations accurately, a large number of Peripheral Blood Mononuclear Cells (PBMCs) must be obtained from blood samples. Traditional manual isolation and SepMateTM isolation of PBMCs consistently yield blood-stained plasma layers and overall low numbers of CD4+ and CD8+ cells. Here, we describe an optimized protocol, using PBS with EDTA to increase PBMC yield from pregnant patients. This protocol enables analysis of CD4+, CD8+, and Regulatory T Cells and is potentially applicable to any immune cell population.
For complete details on the use and execution of this protocol, please refer to the SepMateTM website https://www.stemcell.com/products/brands/SepMateTM-pbmc-isolation.html.