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Ficoll-Paque Pharmacia mononuclear cells mncs
Mononuclear Cells Mncs, supplied by Ficoll-Paque Pharmacia, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 92 stars, based on 4 article reviews
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mononuclear cells mncs - by Bioz Stars, 2020-09
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Isolation:

Article Title: SARS-CoV-2 Entry Receptor ACE2 Is Expressed on Very Small CD45− Precursors of Hematopoietic and Endothelial Cells and in Response to Virus Spike Protein Activates the Nlrp3 Inflammasome
Article Snippet: .. Briefly, mononuclear cells (MNCs) were isolated from hUCB samples by density-gradient centrifugation on a Ficoll–Paque gradient (ρ = 1.077 g/mL; GE Healthcare). .. MNCs were then labelled with anti-CD34 or anti-CD133 MicroBeads (Miltenyi Biotec) and separated on magnetic columns (Miltenyi Biotec).

Article Title: Dose-Dependent Circulating Immunoglobulin A Antibody-Secreting Cell and Serum Antibody Responses in Swedish Volunteers to an Oral Inactivated Enterotoxigenic Escherichia coli Vaccine
Article Snippet: .. Mononuclear cells (MNCs) from heparinized venous blood were isolated by standard gradient centrifugation on Ficoll-Paque (Pharmacia Biotech AB, Uppsala, Sweden). .. The numbers of IgA-secreting cells and antigen-specific IgA ASCs were measured by a micromodification of the original enzyme-linked immunospot assay ( , ).

Article Title: Chemical, Physical and Biological Triggers of Evolutionary Conserved Bcl-xL-Mediated Apoptosis
Article Snippet: .. Mononuclear cells (MNCs) were isolated by density gradient separation (Ficoll-Paque PREMIUM; GE Healthcare, Little Chalfont, Buckinghamshire, UK) and immediately analyzed or vitally frozen for later use. .. Cells were maintained in mononuclear cell medium (MCM; Promocell, Heidelberg, Germany) or in a 25% HS-5 CM plus 75% RPMI 1640 medium mix.

Article Title: Clonal expansion is a characteristic feature of the B-cell repertoire of patients with rheumatoid arthritis
Article Snippet: .. Mononuclear cells (MNCs) were isolated from cell suspensions from blood, synovial fluid, and synovial tissue by density gradient centrifugation (Ficoll-Paque; Pharmacia LKB Biotechnology, Piscataway, NJ, USA). .. Isolation of DNA and RNA, and preparation of complementary DNA Genomic DNA was isolated from MNCs using the Puregene DNA Isolation Kit (Gentra Systems, Mineapolis, MN, USA) and total RNA was isolated using Ultraspec RNA (Biotech Laboratories, Houston, TX, USA).

Article Title: Chemical, physical and biological triggers of evolutionary conserved Bcl-xL-mediated apoptosis
Article Snippet: .. Mononuclear cells (MNCs) were isolated by density gradient separation (Ficoll-Paque PREMIUM; GE Healthcare, Little Chalfont, Buckinghamshire, UK) and immediately analyzed or vitally frozen for later use. .. Cells were maintained in mononuclear cell medium (MCM; Promocell, Heidelberg, Germany) or in a 25% HS-5 CM plus 75% RPMI 1640 medium mix.

Article Title: Endothelial progenitor cells with stem cells enhance osteogenic efficacy
Article Snippet: .. Mononuclear cells (MNCs) were isolated from all heparinized BM aspirates by density gradient centrifugation using Ficoll Paque Plus (GE Healthcare, Uppsala, Sweden). .. The MNCs were then washed in PBS and cultured in culture flasks (Corning, New York, USA) with α-MEM (HyClone, Logan, USA) containing 10% (V/V) fetal bovine serum (GIBCO, Carlsbad, USA).

Centrifugation:

Article Title: SARS-CoV-2 Entry Receptor ACE2 Is Expressed on Very Small CD45− Precursors of Hematopoietic and Endothelial Cells and in Response to Virus Spike Protein Activates the Nlrp3 Inflammasome
Article Snippet: .. Briefly, mononuclear cells (MNCs) were isolated from hUCB samples by density-gradient centrifugation on a Ficoll–Paque gradient (ρ = 1.077 g/mL; GE Healthcare). .. MNCs were then labelled with anti-CD34 or anti-CD133 MicroBeads (Miltenyi Biotec) and separated on magnetic columns (Miltenyi Biotec).

Gradient Centrifugation:

Article Title: Dose-Dependent Circulating Immunoglobulin A Antibody-Secreting Cell and Serum Antibody Responses in Swedish Volunteers to an Oral Inactivated Enterotoxigenic Escherichia coli Vaccine
Article Snippet: .. Mononuclear cells (MNCs) from heparinized venous blood were isolated by standard gradient centrifugation on Ficoll-Paque (Pharmacia Biotech AB, Uppsala, Sweden). .. The numbers of IgA-secreting cells and antigen-specific IgA ASCs were measured by a micromodification of the original enzyme-linked immunospot assay ( , ).

Article Title: Clonal expansion is a characteristic feature of the B-cell repertoire of patients with rheumatoid arthritis
Article Snippet: .. Mononuclear cells (MNCs) were isolated from cell suspensions from blood, synovial fluid, and synovial tissue by density gradient centrifugation (Ficoll-Paque; Pharmacia LKB Biotechnology, Piscataway, NJ, USA). .. Isolation of DNA and RNA, and preparation of complementary DNA Genomic DNA was isolated from MNCs using the Puregene DNA Isolation Kit (Gentra Systems, Mineapolis, MN, USA) and total RNA was isolated using Ultraspec RNA (Biotech Laboratories, Houston, TX, USA).

Article Title: Endothelial progenitor cells with stem cells enhance osteogenic efficacy
Article Snippet: .. Mononuclear cells (MNCs) were isolated from all heparinized BM aspirates by density gradient centrifugation using Ficoll Paque Plus (GE Healthcare, Uppsala, Sweden). .. The MNCs were then washed in PBS and cultured in culture flasks (Corning, New York, USA) with α-MEM (HyClone, Logan, USA) containing 10% (V/V) fetal bovine serum (GIBCO, Carlsbad, USA).

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  • 94
    Ficoll-Paque Pharmacia pbmc isolation
    Ex vivo production of IL-1β ( a ) and IFN-γ ( b ) by <t>PBMCs</t> stimulated with LPS, heat-killed S. aureus ( S.a ), heat-killed C. albicans ( C.a ), and sonicated M. tuberculosis ( M.tb ) at baseline, and at 1 and 4 days after vaccination with TFV or BCG vaccine. Wilcoxon signed-rank test comparing values before and after vaccination with the same stimulus; N = 29, ^ p = 0.06, * p
    Pbmc Isolation, supplied by Ficoll-Paque Pharmacia, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmc isolation/product/Ficoll-Paque Pharmacia
    Average 94 stars, based on 1 article reviews
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    92
    Ficoll-Paque Pharmacia pbmcs
    Platelets Are Critical for the Production of IL-1 Cytokines by Human Primary Monocytes (A) Representative FACS dot plots of CD41 and <t>CD14</t> expression in human <t>PBMCs,</t> standard (Std-Mo), or platelet-depleted (PD-Mo) CD14 + monocytes (see STAR Methods ). (B) Quantification of platelets (CD41 + CD14 − ), platelet-monocyte aggregates (CD41 + CD14 + ), and platelet-free monocytes (CD41 − CD14 + ) in PBMCs and isolated monocytes. (C) IL-1β and TNF-α levels in cell-free supernatants of Std-Mo versus PD-Mo, or in platelet-depleted monocytes replenished with autologous platelets (PD-Mo + Plts, 50:1 Ptl:Cell), and stimulated as indicated. (D and E) IL-1β levels released by non-canonically activated Std-Mo, PD-Mo, or PD-Mo + Plts, stimulated with LPS (1 μg mL −1 ) for 16 h (D) or by inflammasome-activated THP-1 s ± platelets (E). Data is presented as floating bars (with mean and minimum to maximum values) and pooled from independent experiments. Each symbol represents the average from technical triplicates per blood donor. Data in (D) shows bar graph with Mean + SD, from a representative of two independent experiments. See also Figure S2 .
    Pbmcs, supplied by Ficoll-Paque Pharmacia, used in various techniques. Bioz Stars score: 92/100, based on 120 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Ficoll-Paque Pharmacia orov infection pbmc
    <t>OROV</t> infection of <t>PBMCs</t> induces the expression of cytokines and innate immune response genes. Cell lysates from PBMCs obtained from healthy donors ( n = 3) and infected with OROV at MOI 10 were processed for RNA extraction and RT-PCR. ( A ) Scheme of the RNA PAMPs recognition pathways analyzed. ( B – E ) SYBR Green qRT-PCR data analysis of gene expression are shown in fold change (2^∆∆ C T). Graphics show results from two independent experiments. Bars represent mean ± SEM. All times post-infection were compared to the time 0 h samples (noninfected) by two-way ANOVA and Dunnett’s multiple comparisons test. **** p -value
    Orov Infection Pbmc, supplied by Ficoll-Paque Pharmacia, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Ex vivo production of IL-1β ( a ) and IFN-γ ( b ) by PBMCs stimulated with LPS, heat-killed S. aureus ( S.a ), heat-killed C. albicans ( C.a ), and sonicated M. tuberculosis ( M.tb ) at baseline, and at 1 and 4 days after vaccination with TFV or BCG vaccine. Wilcoxon signed-rank test comparing values before and after vaccination with the same stimulus; N = 29, ^ p = 0.06, * p

    Journal: European Journal of Clinical Microbiology & Infectious Diseases

    Article Title: Differential effects of BCG vaccine on immune responses induced by vi polysaccharide typhoid fever vaccination: an explorative randomized trial

    doi: 10.1007/s10096-020-03813-y

    Figure Lengend Snippet: Ex vivo production of IL-1β ( a ) and IFN-γ ( b ) by PBMCs stimulated with LPS, heat-killed S. aureus ( S.a ), heat-killed C. albicans ( C.a ), and sonicated M. tuberculosis ( M.tb ) at baseline, and at 1 and 4 days after vaccination with TFV or BCG vaccine. Wilcoxon signed-rank test comparing values before and after vaccination with the same stimulus; N = 29, ^ p = 0.06, * p

    Article Snippet: PBMC isolation and stimulation PBMCs were isolated using density-gradient separation over Ficoll-Paque (GE Healthcare, UK ).

    Techniques: Ex Vivo, Sonication

    Ex vivo production of IL-6 ( a ), IL-10 ( b ), IFN-γ ( c ), and IL-22 ( d ) by PBMCs stimulated with LPS, heat-killed S. aureus ( S.a ), heat-killed C. albicans ( C.a ), and sonicated M. tuberculosis ( M.tb ), before TFV or BCG vaccination (baseline) and at 2 weeks and 3 months after TFV vaccination. Wilcoxon signed-rank test comparing values before and after vaccination with the same stimulus; N = 29, * p

    Journal: European Journal of Clinical Microbiology & Infectious Diseases

    Article Title: Differential effects of BCG vaccine on immune responses induced by vi polysaccharide typhoid fever vaccination: an explorative randomized trial

    doi: 10.1007/s10096-020-03813-y

    Figure Lengend Snippet: Ex vivo production of IL-6 ( a ), IL-10 ( b ), IFN-γ ( c ), and IL-22 ( d ) by PBMCs stimulated with LPS, heat-killed S. aureus ( S.a ), heat-killed C. albicans ( C.a ), and sonicated M. tuberculosis ( M.tb ), before TFV or BCG vaccination (baseline) and at 2 weeks and 3 months after TFV vaccination. Wilcoxon signed-rank test comparing values before and after vaccination with the same stimulus; N = 29, * p

    Article Snippet: PBMC isolation and stimulation PBMCs were isolated using density-gradient separation over Ficoll-Paque (GE Healthcare, UK ).

    Techniques: Ex Vivo, Sonication

    Platelets Are Critical for the Production of IL-1 Cytokines by Human Primary Monocytes (A) Representative FACS dot plots of CD41 and CD14 expression in human PBMCs, standard (Std-Mo), or platelet-depleted (PD-Mo) CD14 + monocytes (see STAR Methods ). (B) Quantification of platelets (CD41 + CD14 − ), platelet-monocyte aggregates (CD41 + CD14 + ), and platelet-free monocytes (CD41 − CD14 + ) in PBMCs and isolated monocytes. (C) IL-1β and TNF-α levels in cell-free supernatants of Std-Mo versus PD-Mo, or in platelet-depleted monocytes replenished with autologous platelets (PD-Mo + Plts, 50:1 Ptl:Cell), and stimulated as indicated. (D and E) IL-1β levels released by non-canonically activated Std-Mo, PD-Mo, or PD-Mo + Plts, stimulated with LPS (1 μg mL −1 ) for 16 h (D) or by inflammasome-activated THP-1 s ± platelets (E). Data is presented as floating bars (with mean and minimum to maximum values) and pooled from independent experiments. Each symbol represents the average from technical triplicates per blood donor. Data in (D) shows bar graph with Mean + SD, from a representative of two independent experiments. See also Figure S2 .

    Journal: Cell Reports

    Article Title: Platelets Fuel the Inflammasome Activation of Innate Immune Cells

    doi: 10.1016/j.celrep.2020.107615

    Figure Lengend Snippet: Platelets Are Critical for the Production of IL-1 Cytokines by Human Primary Monocytes (A) Representative FACS dot plots of CD41 and CD14 expression in human PBMCs, standard (Std-Mo), or platelet-depleted (PD-Mo) CD14 + monocytes (see STAR Methods ). (B) Quantification of platelets (CD41 + CD14 − ), platelet-monocyte aggregates (CD41 + CD14 + ), and platelet-free monocytes (CD41 − CD14 + ) in PBMCs and isolated monocytes. (C) IL-1β and TNF-α levels in cell-free supernatants of Std-Mo versus PD-Mo, or in platelet-depleted monocytes replenished with autologous platelets (PD-Mo + Plts, 50:1 Ptl:Cell), and stimulated as indicated. (D and E) IL-1β levels released by non-canonically activated Std-Mo, PD-Mo, or PD-Mo + Plts, stimulated with LPS (1 μg mL −1 ) for 16 h (D) or by inflammasome-activated THP-1 s ± platelets (E). Data is presented as floating bars (with mean and minimum to maximum values) and pooled from independent experiments. Each symbol represents the average from technical triplicates per blood donor. Data in (D) shows bar graph with Mean + SD, from a representative of two independent experiments. See also Figure S2 .

    Article Snippet: Isolation of CD14+ human Monocytes Venous blood was collected in S-Monovette® K3EDTA tubes and PBMCs were obtained by density gradient centrifugation in Ficoll-Paque PLUS.

    Techniques: FACS, Expressing, Isolation

    Unstimulated neutrophils display a spectrum of densities . Neutrophils were first isolated using Ficoll density centrifugation. Thereafter isolated neutrophils were centrifuged on top of Percoll with different densities. ( A ) Depicted is the percentage of total neutrophils in the “PBMC layer” after density centrifugation with different densities of Percoll ( n = 9). ( B ) The lag time in which higher density neutrophils (HDNs) and lower density neutrophils (LDNs) contain growth of GFP labeled S. Aureus ( n = 7). ( C ) The percentage of the original population of lymphocytes that divided after stimulation in the absence of neutrophils, in the presence of all neutrophils and in the presence of either LDN or HDN of the same donor ( n = 7). ( D ) Division index (average number of cell divisions that a cell in the original population has undergone) of lymphocytes under the same conditions as ( C ). ( E ) Proliferation index (the total number of divisions divided by the number of cells that went into division) of lymphocytes under the same conditions as ( C ). For all graphs median ± IQR% is shown. Data are analyzed using Friedman test without correction for multiple comparisons. ( C‐E ) all conditions were tested, but only statistically significant results are indicated

    Journal: Journal of Leukocyte Biology

    Article Title: On the origin of low‐density neutrophils, et al. On the origin of low‐density neutrophils

    doi: 10.1002/JLB.5HR0120-459R

    Figure Lengend Snippet: Unstimulated neutrophils display a spectrum of densities . Neutrophils were first isolated using Ficoll density centrifugation. Thereafter isolated neutrophils were centrifuged on top of Percoll with different densities. ( A ) Depicted is the percentage of total neutrophils in the “PBMC layer” after density centrifugation with different densities of Percoll ( n = 9). ( B ) The lag time in which higher density neutrophils (HDNs) and lower density neutrophils (LDNs) contain growth of GFP labeled S. Aureus ( n = 7). ( C ) The percentage of the original population of lymphocytes that divided after stimulation in the absence of neutrophils, in the presence of all neutrophils and in the presence of either LDN or HDN of the same donor ( n = 7). ( D ) Division index (average number of cell divisions that a cell in the original population has undergone) of lymphocytes under the same conditions as ( C ). ( E ) Proliferation index (the total number of divisions divided by the number of cells that went into division) of lymphocytes under the same conditions as ( C ). For all graphs median ± IQR% is shown. Data are analyzed using Friedman test without correction for multiple comparisons. ( C‐E ) all conditions were tested, but only statistically significant results are indicated

    Article Snippet: After phenotyping the lower and normal density neutrophils, a difference in numbers of CD16dim /CD62Lhigh neutrophils between the PBMC and the granulocyte fraction after Ficoll‐Paque centrifugation (median of 42.0% in LDNs vs. 11.9% in NDNs) was again apparent (Fig. ).

    Techniques: Isolation, Centrifugation, Labeling

    OROV infection of PBMCs induces the expression of cytokines and innate immune response genes. Cell lysates from PBMCs obtained from healthy donors ( n = 3) and infected with OROV at MOI 10 were processed for RNA extraction and RT-PCR. ( A ) Scheme of the RNA PAMPs recognition pathways analyzed. ( B – E ) SYBR Green qRT-PCR data analysis of gene expression are shown in fold change (2^∆∆ C T). Graphics show results from two independent experiments. Bars represent mean ± SEM. All times post-infection were compared to the time 0 h samples (noninfected) by two-way ANOVA and Dunnett’s multiple comparisons test. **** p -value

    Journal: Viruses

    Article Title: Oropouche Virus Infects, Persists and Induces IFN Response in Human Peripheral Blood Mononuclear Cells as Identified by RNA PrimeFlow™ and qRT-PCR Assays

    doi: 10.3390/v12070785

    Figure Lengend Snippet: OROV infection of PBMCs induces the expression of cytokines and innate immune response genes. Cell lysates from PBMCs obtained from healthy donors ( n = 3) and infected with OROV at MOI 10 were processed for RNA extraction and RT-PCR. ( A ) Scheme of the RNA PAMPs recognition pathways analyzed. ( B – E ) SYBR Green qRT-PCR data analysis of gene expression are shown in fold change (2^∆∆ C T). Graphics show results from two independent experiments. Bars represent mean ± SEM. All times post-infection were compared to the time 0 h samples (noninfected) by two-way ANOVA and Dunnett’s multiple comparisons test. **** p -value

    Article Snippet: Human PBMC Culture and OROV Infection PBMC were obtained by gradient centrifugation with Ficoll-Paque 1.077 g/mL, according to the manufacturer’s instructions with some modifications.

    Techniques: Infection, Expressing, RNA Extraction, Reverse Transcription Polymerase Chain Reaction, SYBR Green Assay, Quantitative RT-PCR

    OROV infection and replication in PBMCs is favored when the type I IFN receptor is blocked and when the cells are treated with Dexamethasone. PBMC were pretreated for 2 h with Dexamethasone (Dexo) 1 µM and human anti-IFNAR antibody 5 ng/mL, and infected with OROV at MOI 1. Cell lysates and supernatants were collected for analysis. ( A ) qRT-PCR of cell lysate and supernatant ( B ) for OROV RNA detection through the time of infection. Data were compared by two-way ANOVA and Tukey’s multiple comparison test. ( C ) Infectious particles released in the supernatant were measured by FFA, 48 hpi. Treatment groups were compared by one-way ANOVA and Dunn’s multiple comparisons test. Bars represent viral load ± SEM. **** p -value

    Journal: Viruses

    Article Title: Oropouche Virus Infects, Persists and Induces IFN Response in Human Peripheral Blood Mononuclear Cells as Identified by RNA PrimeFlow™ and qRT-PCR Assays

    doi: 10.3390/v12070785

    Figure Lengend Snippet: OROV infection and replication in PBMCs is favored when the type I IFN receptor is blocked and when the cells are treated with Dexamethasone. PBMC were pretreated for 2 h with Dexamethasone (Dexo) 1 µM and human anti-IFNAR antibody 5 ng/mL, and infected with OROV at MOI 1. Cell lysates and supernatants were collected for analysis. ( A ) qRT-PCR of cell lysate and supernatant ( B ) for OROV RNA detection through the time of infection. Data were compared by two-way ANOVA and Tukey’s multiple comparison test. ( C ) Infectious particles released in the supernatant were measured by FFA, 48 hpi. Treatment groups were compared by one-way ANOVA and Dunn’s multiple comparisons test. Bars represent viral load ± SEM. **** p -value

    Article Snippet: Human PBMC Culture and OROV Infection PBMC were obtained by gradient centrifugation with Ficoll-Paque 1.077 g/mL, according to the manufacturer’s instructions with some modifications.

    Techniques: Infection, Quantitative RT-PCR, RNA Detection

    Human peripheral blood monocytes and lymphocytes are susceptible to OROV infection but generate low yields of infectious particles. Human peripheral blood mononuclear cells (PBMCs) were obtained from whole blood of healthy donors, infected in vitro and analyzed by different methodologies. ( A ) PBMCs were seeded in 24-well plates and infected with OROV MOI 0.1; 1 and 10. Cell lysates and supernatants were collected to RNA quantification by qRT-PCR ( n = 3). ( B ) The supernatants were also analyzed by FFA assay. Symbols represent the mean of viral load ± SEM. ( C ) PBMCs from healthy donors ( n = 2) were infected with MOI 1, submitted RNA PrimeFlow™ protocol 24 hpi and flow cytometry. CD3 + and CD14 + HLA DR + percentage of events with OROV Grna, and the gating strategy are shown. ( D ) Detection of OROV 48 hpi in cells infected with MOI 2, by confocal microscopy. OROV proteins in red (Alexa Fluor 594); genome (gRNA) in magenta (AlexaFluor 647); antigenome (agRNA) in green (AlexaFluor 488); DAPI (blue). Images with 63x times magnification. Scales at 25 μm.

    Journal: Viruses

    Article Title: Oropouche Virus Infects, Persists and Induces IFN Response in Human Peripheral Blood Mononuclear Cells as Identified by RNA PrimeFlow™ and qRT-PCR Assays

    doi: 10.3390/v12070785

    Figure Lengend Snippet: Human peripheral blood monocytes and lymphocytes are susceptible to OROV infection but generate low yields of infectious particles. Human peripheral blood mononuclear cells (PBMCs) were obtained from whole blood of healthy donors, infected in vitro and analyzed by different methodologies. ( A ) PBMCs were seeded in 24-well plates and infected with OROV MOI 0.1; 1 and 10. Cell lysates and supernatants were collected to RNA quantification by qRT-PCR ( n = 3). ( B ) The supernatants were also analyzed by FFA assay. Symbols represent the mean of viral load ± SEM. ( C ) PBMCs from healthy donors ( n = 2) were infected with MOI 1, submitted RNA PrimeFlow™ protocol 24 hpi and flow cytometry. CD3 + and CD14 + HLA DR + percentage of events with OROV Grna, and the gating strategy are shown. ( D ) Detection of OROV 48 hpi in cells infected with MOI 2, by confocal microscopy. OROV proteins in red (Alexa Fluor 594); genome (gRNA) in magenta (AlexaFluor 647); antigenome (agRNA) in green (AlexaFluor 488); DAPI (blue). Images with 63x times magnification. Scales at 25 μm.

    Article Snippet: Human PBMC Culture and OROV Infection PBMC were obtained by gradient centrifugation with Ficoll-Paque 1.077 g/mL, according to the manufacturer’s instructions with some modifications.

    Techniques: Infection, In Vitro, Quantitative RT-PCR, Flow Cytometry, Confocal Microscopy