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. Author manuscript; available in PMC: 2016 May 1.
Published in final edited form as: Cell Immunol. 2015 Feb 14;295(1):19–28. doi: 10.1016/j.cellimm.2015.02.006

Fig. 1. Optimization of cynomolgus PBMC isolation for subsequent Treg sorting.

Fig. 1

(A) Outlines of three methods tested to isolate cynomolgus PBMC. (B) Percentages of monocytes, B cells, and T cells in PBMC isolates and percentages of Treg in total CD4+ T cells. Lymphocytes were gated based on forward (FSC) and side-scatter (SSC) of singlets. Monocytes were gated on CD14+; B cells on CD3−CD20+; T cells on CD3+CD20−; and Treg on CD25+CD127−/loCD3+CD4+ T cells. Data are representative of 3 individual experiments. (C) Percent recovery of monocytes, B cells, CD4+ T cells and Treg from equivalent amounts of cynomolgus blood using different PBMC isolation methods. Absolute counts per ml blood of each type of cell were determined by counting beads as: number of cell events / number of bead events X total number of bead added / volume of sample. Percent recovery was defined as: absolute number of cells in PBMC isolated from 1ml blood / absolute number of cells in 1ml blood treated with red blood cell lysis buffer X 100%. Data are means + 1SD from 3 individual experiments. (D) Absolute number of indicated cells in PBMC isolated using the three methods. Whole blood was used as control. Data are means + 1SD from 3 individual experiments.