blood mononuclear cells pbmcs  (Becton Dickinson)

Journal: Immunologic Research

Article Title: Impaired natural killer cell subset phenotypes in human obesity

doi: 10.1007/s12026-018-8989-4

Figure Lengend Snippet: Flow cytometric analyses of interferon gamma (IFN-γ) expression on NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of IFN-γ expression in CD56 bright and CD56 dim NK cells of a normal-weight and an obese individual. c Frequency of IFN-γ-expressing NK cells in PBMCs. d , e IFN-γ expression in CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

Article Snippet: For flow cytometric analyses, PBMCs were stained with the following antibodies (all from BD Biosciences, San Diego, USA, unless otherwise indicated): CD3 conjugated with phycoerythrin (PE)-Cy 7, CD4 conjugated with allophycocyanin (APC), CD 8 conjugated with PE, CD14 conjugated with fluorescein isothiocyanate (FITC), CD20 conjugated with APC-H7, CD56 conjugated with APC, CD253 (TRAIL; tumor necrosis factor-related apoptosis-inducing ligand) conjugated with PE, Ob-R (leptin receptor) conjugated with fluorescein (R & D Systems, Minneapolis, MN, USA), CD314 (NKG2D) conjugated with PE-CF594, CD25 conjugated with PE CF594, CD69 conjugated with FITC, and CD107a conjugated with PE.

Techniques: Flow Cytometry, Expressing, Isolation, FACS

Journal: Immunologic Research

Article Title: Impaired natural killer cell subset phenotypes in human obesity

doi: 10.1007/s12026-018-8989-4

Figure Lengend Snippet: Flow cytometric analyses of NK cells and NK cell subsets in PBMCs (peripheral blood mononuclear cells) isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of CD56 bright and CD56 dim NK cells of a normal-weight and an obese subject. c Frequency of NK cells in PBMCs. d , e Expression of CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

Article Snippet: For flow cytometric analyses, PBMCs were stained with the following antibodies (all from BD Biosciences, San Diego, USA, unless otherwise indicated): CD3 conjugated with phycoerythrin (PE)-Cy 7, CD4 conjugated with allophycocyanin (APC), CD 8 conjugated with PE, CD14 conjugated with fluorescein isothiocyanate (FITC), CD20 conjugated with APC-H7, CD56 conjugated with APC, CD253 (TRAIL; tumor necrosis factor-related apoptosis-inducing ligand) conjugated with PE, Ob-R (leptin receptor) conjugated with fluorescein (R & D Systems, Minneapolis, MN, USA), CD314 (NKG2D) conjugated with PE-CF594, CD25 conjugated with PE CF594, CD69 conjugated with FITC, and CD107a conjugated with PE.

Techniques: Flow Cytometry, Isolation, FACS, Expressing

Journal: Immunologic Research

Article Title: Impaired natural killer cell subset phenotypes in human obesity

doi: 10.1007/s12026-018-8989-4

Figure Lengend Snippet: Flow cytometric analyses of NKG2D receptor expression on NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of NKG2D expression in CD56 bright and CD56 dim NK cells of a normal-weight and an obese subject. c Frequency of NKG2D-positive NK cells in PBMCs. d , e Expression of NKG2D in CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

Article Snippet: For flow cytometric analyses, PBMCs were stained with the following antibodies (all from BD Biosciences, San Diego, USA, unless otherwise indicated): CD3 conjugated with phycoerythrin (PE)-Cy 7, CD4 conjugated with allophycocyanin (APC), CD 8 conjugated with PE, CD14 conjugated with fluorescein isothiocyanate (FITC), CD20 conjugated with APC-H7, CD56 conjugated with APC, CD253 (TRAIL; tumor necrosis factor-related apoptosis-inducing ligand) conjugated with PE, Ob-R (leptin receptor) conjugated with fluorescein (R & D Systems, Minneapolis, MN, USA), CD314 (NKG2D) conjugated with PE-CF594, CD25 conjugated with PE CF594, CD69 conjugated with FITC, and CD107a conjugated with PE.

Techniques: Flow Cytometry, Expressing, Isolation, FACS

Journal: Immunologic Research

Article Title: Impaired natural killer cell subset phenotypes in human obesity

doi: 10.1007/s12026-018-8989-4

Figure Lengend Snippet: Flow cytometric analyses of leptin receptor (Ob-R) expression in NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a Frequency of Ob-R expressing NK cells in PBMCs. b , c Expression of Ob-R in CD56 bright ( b ) and CD56 dim ( c ) NK cells. Data are expressed as mean ± SEM

Article Snippet: For flow cytometric analyses, PBMCs were stained with the following antibodies (all from BD Biosciences, San Diego, USA, unless otherwise indicated): CD3 conjugated with phycoerythrin (PE)-Cy 7, CD4 conjugated with allophycocyanin (APC), CD 8 conjugated with PE, CD14 conjugated with fluorescein isothiocyanate (FITC), CD20 conjugated with APC-H7, CD56 conjugated with APC, CD253 (TRAIL; tumor necrosis factor-related apoptosis-inducing ligand) conjugated with PE, Ob-R (leptin receptor) conjugated with fluorescein (R & D Systems, Minneapolis, MN, USA), CD314 (NKG2D) conjugated with PE-CF594, CD25 conjugated with PE CF594, CD69 conjugated with FITC, and CD107a conjugated with PE.

Techniques: Flow Cytometry, Expressing, Isolation

Journal: Immunologic Research

Article Title: Impaired natural killer cell subset phenotypes in human obesity

doi: 10.1007/s12026-018-8989-4

Figure Lengend Snippet: Flow cytometric analyses of CD69 expression on NK cells and NK cell subsets in PBMCs isolated from normal-weight (nw) and obese (ob) individuals. a , b Exemplary FACS plots of CD69 expression in CD56 bright and CD56 dim NK cells of a normal-weight and an obese subject. c Frequency of CD69-positive NK cells in PBMCs. d , e Expression of CD69 in CD56 bright ( d ) and CD56 dim ( e ) NK cells. Data are expressed as mean ± SEM. * P

Article Snippet: For flow cytometric analyses, PBMCs were stained with the following antibodies (all from BD Biosciences, San Diego, USA, unless otherwise indicated): CD3 conjugated with phycoerythrin (PE)-Cy 7, CD4 conjugated with allophycocyanin (APC), CD 8 conjugated with PE, CD14 conjugated with fluorescein isothiocyanate (FITC), CD20 conjugated with APC-H7, CD56 conjugated with APC, CD253 (TRAIL; tumor necrosis factor-related apoptosis-inducing ligand) conjugated with PE, Ob-R (leptin receptor) conjugated with fluorescein (R & D Systems, Minneapolis, MN, USA), CD314 (NKG2D) conjugated with PE-CF594, CD25 conjugated with PE CF594, CD69 conjugated with FITC, and CD107a conjugated with PE.

Techniques: Flow Cytometry, Expressing, Isolation, FACS

Journal: The Journal of Experimental Medicine

Article Title: Low-dose irradiation promotes tissue revascularization through VEGF release from mast cells and MMP-9-mediated progenitor cell mobilization

doi: 10.1084/jem.20050959

Figure Lengend Snippet: Irradiation-induced angiogenesis is dictated by MMP-9–mediated factor release from mast and stroma cells and incorporation of BM-derived cells. (A–C) HL ischemia was induced in Sl/Sl d and WBB6F1 +/+ mice, followed by 2 Gy IR or no IR ( n = 7). PBMCs were analyzed for the presence of CFU-Cs (A) and CFU-ECs (B). (C) Plasma VEGF was determined ( n = 7/group). (D) HL ischemia was induced in MMP-9 +/+ mice followed by 2 Gy IR. Mice were subdivided into groups receiving neutralizing mAbs against VEGFR-1, VEGFR-2 or a combination of both mAbs. (a) Blood flow was determined. Insert: Percentage of mice showing limb rescue after ligation. (b) Macroscopic pictures taken of ischemic and contralateral limb 7 d after ligation (*P

Article Snippet: To understand which cell types increase after IR, we analyzed peripheral blood mononuclear cells (PBMCs) via FACS (Becton Dickinson) using mAbs against c-kit and VEGFR-2 after IR.

Techniques: Irradiation, Derivative Assay, Mouse Assay, Flow Cytometry, Ligation

Journal: Frontiers in Immunology

Article Title: T Cell Phenotype and T Cell Receptor Repertoire in Patients with Major Depressive Disorder

doi: 10.3389/fimmu.2018.00291

Figure Lengend Snippet: CXCR3 and CCR6 expression in T cells of major depressive disorder (MDD) patients and non-depressed controls. (A) CXCR3-expressing T cells were identified by flow cytometric analysis of peripheral blood mononuclear cells from MDD patients and matched non-depressed controls (CTR). Displayed values are frequencies of CXCR3 + T cells expressed as a percentage of live CD3 + lymphocytes from a representative case–control pair. (B) Percentages of CXCR3-expressing total T cells, CD4 + , and CD8 + T cells were quantified in our cohort ( n = 40). (C,D) Similar analyses were conducted for the surface expression of CCR6 on total T cells as well as on the CD4 + and CD8 + T cell subsets. (E) The CXCR3 ligands CXCL10 and CXCL11 were quantified in sera of MDD patients and matched controls using a cytometric bead array ( n = 38). (F) Surface CD3 MFI levels were measured by flow cytometric analysis of CD4 + and CD8 + T cells from MDD patients and matched controls ( n = 40). All graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used. SSC-A, side scatter-area; CXCR3, CXC-chemokine receptor type 3; CCR6, CC-chemokine receptor type 6; MFI, median fluorescence intensity.

Article Snippet: Cell Purification For qRT-PCR and TCR sequencing analyses, CD4+ T cells were purified using magnetic beads (negative selection by BD IMag Human CD4 T Lymphocyte Enrichment Set, BD Biosciences) from PBMC aliquots thawed in cell separation buffer (1% human serum, 2 mM EDTA in PBS).

Techniques: Expressing, Flow Cytometry, Fluorescence

Journal: Frontiers in Immunology

Article Title: T Cell Phenotype and T Cell Receptor Repertoire in Patients with Major Depressive Disorder

doi: 10.3389/fimmu.2018.00291

Figure Lengend Snippet: Surface and intracellular staining of CXCR3. (A) Percentages of CXCR3- and CCR6-expressing CD3 − lymphocytes (non-T cells) were quantified in major depressive disorder (MDD) patients and matched non-depressed controls (CTR) ( n = 40). (B) A representative plot shows fluorescence intensity of CXCR3 expression in intact (surface CXCR3; light gray-shaded curve) relative to fixed and permeabilized T cells (total cellular CXCR3; dark gray-shaded curve). Isotype-matched negative controls were used at the same concentration before fixation (black-dashed curve) and after fixation-permeabilization (gray-dashed curve) and showed no positive staining for CXCR3. (C) Total cellular CXCR3 MFI levels were measured by flow cytometric analysis of fixed and permeabilized peripheral blood mononuclear cells (PBMCs) from MDD patients and matched controls ( n = 36). Stained PBMCs were gated on live CD3 + lymphocytes (T cells), CD4 + and CD8 + T cell subsets as well as CD3 − lymphocytes (non-T cells). Graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used. CXCR3: CXC-chemokine receptor type 3; CCR6: CC-chemokine receptor type 6; MFI: median fluorescence intensity.

Article Snippet: Cell Purification For qRT-PCR and TCR sequencing analyses, CD4+ T cells were purified using magnetic beads (negative selection by BD IMag Human CD4 T Lymphocyte Enrichment Set, BD Biosciences) from PBMC aliquots thawed in cell separation buffer (1% human serum, 2 mM EDTA in PBS).

Techniques: Staining, Expressing, Fluorescence, Concentration Assay, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: T Cell Phenotype and T Cell Receptor Repertoire in Patients with Major Depressive Disorder

doi: 10.3389/fimmu.2018.00291

Figure Lengend Snippet: Regulatory T cells in major depressive disorder (MDD) patients and non-depressed controls. (A) Regulatory T cells (Tregs) were identified by flow cytometric analysis of peripheral blood mononuclear cells from MDD patients and matched non-depressed controls (CTR). Displayed values are frequencies of Tregs expressed as a percentage of live CD4 + T cells from a representative case–control pair. (B) Differences in Treg frequency are depicted for the entire cohort ( n = 40). (C) Negatively selected CD4 + T cells from a subsample of patients and matched controls ( n = 20) were analyzed for mRNA expression of the T helper-associated transcription factors Forkhead box P3 ( FOXP3 ), T-box 21 ( T-bet ), GATA binding protein 3 ( GATA3 ), and RAR related orphan receptor C ( RORC ), respectively. Expression was normalized to the geometric mean expression of three housekeeping genes ( IPO8, TBP, RPL13A ). (D) The correlation between the expression levels of the gene FOXP3 in purified CD4 + T cells and the frequency of Tregs expressed as a percentage of CD4 + T cells is plotted ( n = 20). Graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used.

Article Snippet: Cell Purification For qRT-PCR and TCR sequencing analyses, CD4+ T cells were purified using magnetic beads (negative selection by BD IMag Human CD4 T Lymphocyte Enrichment Set, BD Biosciences) from PBMC aliquots thawed in cell separation buffer (1% human serum, 2 mM EDTA in PBS).

Techniques: Flow Cytometry, Expressing, Binding Assay, Purification

Journal: Frontiers in Immunology

Article Title: T Cell Phenotype and T Cell Receptor Repertoire in Patients with Major Depressive Disorder

doi: 10.3389/fimmu.2018.00291

Figure Lengend Snippet: Peripheral blood counts and frequencies of major leukocyte subsets. (A) Absolute peripheral blood granulocyte, monocyte, and lymphocyte counts were obtained from major depressive disorder (MDD) patients and matched non-depressed controls (CTR) using a Coulter Ac·T Diff hematology analyzer ( n = 40). (B) Frequencies of total CD3 + lymphocytes (T cells), CD3 − CD56 − CD19 + B cells and CD3 − CD19 − CD20 − CD14 − CD56 + natural killer (NK) cells were obtained by flow cytometric analysis of thawed peripheral blood mononuclear cells. (C) T cells were further discriminated into CD4 + CD8 − and CD8 + CD4 − subsets. (D) Among NK cells, CD56 low CD16 + cytotoxic (NKc) cells and CD56 high CD16 − regulatory (NKreg) cells were also identified. Graphs depict medians with interquartile ranges. For all comparisons, the Wilcoxon signed-rank test was used.

Article Snippet: Cell Purification For qRT-PCR and TCR sequencing analyses, CD4+ T cells were purified using magnetic beads (negative selection by BD IMag Human CD4 T Lymphocyte Enrichment Set, BD Biosciences) from PBMC aliquots thawed in cell separation buffer (1% human serum, 2 mM EDTA in PBS).

Techniques: Flow Cytometry

Journal: BMC Immunology

Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

doi: 10.1186/s12865-015-0113-0

Figure Lengend Snippet: Signal scatter plots comparing the intensities of gene expressions on microarray chips within Ficoll and CPT-derived PBMC and their immune cell subsets for each healthy donor tested. The expression signals of Ficoll-derived cells were plotted against the expression signals of CPT-derived cells for each cell type and donor. A line of perfect correlation is indicated in all plots. Pearson’s correlation coefficients between each pair of samples are shown at the bottom right of each plot. The data showed that the pattern of gene expression was very similar between Ficoll and CPT-derived cell types, as indicated by the high correlation coefficients

Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

Techniques: Microarray, Cycling Probe Technology, Derivative Assay, Expressing

Journal: BMC Immunology

Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

doi: 10.1186/s12865-015-0113-0

Figure Lengend Snippet: Yield and viability of immune cell subsets prepared from PBMC isolated by either Ficoll or CPT protocol. a The mean number of CD19+, CD8+, CD14+, and CD4+ cells positively selected from Ficoll- and CPT-isolated PBMC collected from 6 healthy donors. The horizontal line denotes the means and each symbol represents an individual cell type isolated from Ficoll-PBMC (filled symbols) or CPT-PBMC (empty symbols) from each of 6 donors. b The mean viability of the same cell subsets. No significant differences ( P

Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

Techniques: Isolation, Cycling Probe Technology

Journal: BMC Immunology

Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

doi: 10.1186/s12865-015-0113-0

Figure Lengend Snippet: Principle component analysis of gene expression showing distinct clusters between individual immune cell subsets but not between the same subsets derived from PBMC isolated by CPT or Ficoll protocols. Principle component analysis (PCA) was performed based on expression of all genes from the microarray. Individual immune cell types are represented by different colors: CD19+ B cells, blue; CD8+ T cells, green; CD14+ monocytes, purple; CD4+ T cells, orange; and total PBMC, red. The method used for PBMC isolation is represented by different symbols of differing size: CPT-based procedure by large circles and Ficoll-based procedure by small circles. PC#1, principle component #1; PC#2, principle component #2

Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

Techniques: Expressing, Derivative Assay, Isolation, Cycling Probe Technology, Microarray

Journal: BMC Immunology

Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

doi: 10.1186/s12865-015-0113-0

Figure Lengend Snippet: Yield and quality of RNA recovered from total PBMC and their individual immune cell subsets did not significantly differ between PBMC isolated by Ficoll and CPT methods. a The mean amount of RNA per cell and ( b ) the mean RIN of RNA preparations extracted from total PBMC and their individual immune cell subsets derived from PBMC prepared by Ficoll and CPT methods from 6 health donors. There were no significant differences ( P

Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

Techniques: Isolation, Cycling Probe Technology, Derivative Assay

Journal: BMC Immunology

Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

doi: 10.1186/s12865-015-0113-0

Figure Lengend Snippet: Similar quantities of DNA with comparable quality were obtained from PBMC and immune cell subsets following either Ficoll or CPT protocol of PBMC isolation. a The mean amount of DNA extracted per cell and ( b ) the mean of the 260/280 ratios, as measure of DNA quality, were compared between DNA preparations obtained from total PBMC and immune cell subsets prepared from PBMC isolated by Ficoll and CPT methods from 6 healthy donors. There were no significant differences ( P

Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

Techniques: Cycling Probe Technology, Isolation

Journal: BMC Immunology

Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

doi: 10.1186/s12865-015-0113-0

Figure Lengend Snippet: Schematic outline of the study. PBMC from 6 healthy donors were isolated using Ficoll-Paque gradient fractionation or BD Vacutainer CPT protocol, and cell yield and purity were compared. Subsequently, immune cell subsets were separated by positive selection from PBMC obtained by both methods, and yields and viabilities of the subsets were determined and compared. RNA and DNA were extracted from total PBMC isolated by both protocols and from their subsets, and the nucleic acid yield and quality compared. Finally, gene expression analysis of individual immune cell subsets and total PBMC obtained by Ficoll and CPT isolation methods were performed and the results compared. LN 2 , liquid nitrogen

Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

Techniques: Isolation, Fractionation, Cycling Probe Technology, Selection, Expressing

Journal: BMC Immunology

Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

doi: 10.1186/s12865-015-0113-0

Figure Lengend Snippet: Yield and viability of PBMC isolated by the Ficoll and CPT protocols. a The mean numbers of PBMC per ml of blood obtained by Ficoll or CPT isolation procedure from 6 healthy donors. b The mean viability of PBMC freshly isolated from the same 6 healthy donors by either Ficoll gradient separation or CPT technique. No significant differences ( P

Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

Techniques: Isolation, Cycling Probe Technology

Journal: BMC Immunology

Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

doi: 10.1186/s12865-015-0113-0

Figure Lengend Snippet: Purity of immune cell subsets obtained from PBMC isolated by either Ficoll or CPT procedures. CD19+, CD8+, CD14+, and CD4+ cell subsets were sequentially separated from PBMC isolated by Ficoll and CPT protocols from the same donor and their purity was determined by flow cytometry using antibodies against surface markers specific for individual immune cell types. Filled histograms were given by appropriate immunoglobulin isotype controls. Gates for determining positivity were established using isotype controls so that ~99.0 % of events were negative

Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

Techniques: Isolation, Cycling Probe Technology, Flow Cytometry, Cytometry

Journal: BMC Immunology

Article Title: Immune cell subsets and their gene expression profiles from human PBMC isolated by Vacutainer Cell Preparation Tube (CPT™) and standard density gradient

doi: 10.1186/s12865-015-0113-0

Figure Lengend Snippet: Post-cryopreservation recovery and viability of PBMC isolated by the Ficoll and CPT methods. a The mean percent recovery after cryopreservation of PBMC collected using the Ficoll and CPT protocols. Percent recovery was calculated by dividing the number of PBMC recovered after thawing by the number of cells that were cryopreserved. b The mean viability of recovered PBMC isolated by the Ficoll and CPT technique. No significant differences ( P

Article Snippet: About 45 ml of blood was obtained from each donor using BD Vacutainer tubes containing acid-citrate-dextrose anticoagulant, solution A (ACD-A; BD) from which PBMC were isolated by Ficoll-Paque gradient centrifugation (see below).

Techniques: Isolation, Cycling Probe Technology