Isolation and Analysis of Human Monocyte Subsets

Whole blood was layered onto Ficoll-Paque PLUS (GE Healthcare, Uppsala, Sweden) for density gradient centrifugation and PBMC collection. Cells were immediately stained for flow cytometry, using antibodies to human CD14 (BD, Franklin Lakes, NJ; clone M5E2), CD16 (BD; clone 3G8), and CCR2-APC (R&D Systems, Minneapolis, MN; clone 48607) or isotype matched control antibodies, as described previously.^{12 (link)} Monocyte subsets were acquired with the BD FACSCantoII flow cytometer, and analysis performed using FlowJo software (v. 10.0.8, TreeStar, Ashland, OR). For this analysis, results for CD14+ and CD14+CD16+ expressing monocytes were reported as the percentage of total monocytes; for CCR2+CD14+ and CCR2+CD14+CD16+, as a percentage of CD14+ and CD14+CD16+ monocytes, respectively.

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Morgello S., Buyukturkoglu K., Murray J., Veenstra M., Berman J.W., Byrd D, & Inglese M. (2021). MR spectroscopy and diffusion imaging in people with HIV: relationships to clinical and immunologic findings. Journal of neuroimaging : official journal of the American Society of Neuroimaging, 32(1), 158-170.

Publication 2021

Ficoll-Paque PLUS CCR2-APC

Antibodies Blood Cells Clone Density gradient centrifugation Ficoll Flow cytometry Human Isotype Monocytes

Corresponding Organization :

Other organizations : Icahn School of Medicine at Mount Sinai, Columbia University, Albert Einstein College of Medicine, Queens College, CUNY, The Graduate Center, CUNY, University of Genoa

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Variable analysis

independent variables

Ficoll-Paque PLUS (GE Healthcare, Uppsala, Sweden) for density gradient centrifugation

dependent variables

Percentage of CD14+ and CD14+CD16+ expressing monocytes among total monocytes
Percentage of CCR2+CD14+ and CCR2+CD14+CD16+ monocytes among CD14+ and CD14+CD16+ monocytes, respectively

control variables

Isotype matched control antibodies

controls

Isotype matched control antibodies

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