peripheral blood mononuclear cells pbmcs  (Millipore)

 
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    Name:
    Red Blood Cell Lysis Buffer
    Description:
    Used for both DNA and RNA isolation the buffer is designed for the preferential lysis of red blood cells from human whole blood yielding intact white blood cells free of red blood cells for further applications This buffer is not intended for use with whole blood from any other species
    Catalog Number:
    11814389001
    Price:
    None
    Applications:
    Red blood cell lysis buffer has been used in the isolation of PBMCs (peripheral blood mononuclear cells) from HIV infected patients. It has also been used in the isolation of lung cellsm, naive CD8+ T cells and infected cells from the lung.
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    Structured Review

    Millipore peripheral blood mononuclear cells pbmcs
    Th17 cells increase with reduced Th1 cells in freshly isolated <t>PBMCs</t> and <t>BMMCs</t> from patients with B-ALL. a PBMCs and BMMCs were separated and stimulated with PMA and ionomycin for 5 h in the presence of brefeldin A and subsequently stained with antibodies against CD3, CD8, intracellular IL-17A or IFN-γ. Flow cytometric analysis was used to determine the Th17 and Th1 cell frequencies. Representative dot plots using matching peripheral blood and bone marrow samples from B-ALL patients and healthy donors (HD) were shown. b Statistical data for frequencies of Th17 and Th1 cells within CD4 + T population were shown. c Total RNA was extracted from CD4 + T cells isolated from B-ALL patients and HDs and reverse transcribed into cDNA and subsequently determined for IL-17A and IFN-γ mRNA expression using quantitative PCR. d The frequencies of Th17 cells were significantly decreased in BM when B-ALL patients achieved complete remission (CR). e CD4 + T cells were cultured with or without Nalm-6 cells for 14 days in the presence of OKT3 plus IL-2 (300 units/ml). Then, frequencies of Th17 cells were determined after stimulation with PMA plus ionomycin
    Used for both DNA and RNA isolation the buffer is designed for the preferential lysis of red blood cells from human whole blood yielding intact white blood cells free of red blood cells for further applications This buffer is not intended for use with whole blood from any other species
    https://www.bioz.com/result/peripheral blood mononuclear cells pbmcs/product/Millipore
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2021-03
    86/100 stars

    Images

    1) Product Images from "Increased Th17 cells and IL-17A exist in patients with B cell acute lymphoblastic leukemia and promote proliferation and resistance to daunorubicin through activation of Akt signaling"

    Article Title: Increased Th17 cells and IL-17A exist in patients with B cell acute lymphoblastic leukemia and promote proliferation and resistance to daunorubicin through activation of Akt signaling

    Journal: Journal of Translational Medicine

    doi: 10.1186/s12967-016-0894-9

    Th17 cells increase with reduced Th1 cells in freshly isolated PBMCs and BMMCs from patients with B-ALL. a PBMCs and BMMCs were separated and stimulated with PMA and ionomycin for 5 h in the presence of brefeldin A and subsequently stained with antibodies against CD3, CD8, intracellular IL-17A or IFN-γ. Flow cytometric analysis was used to determine the Th17 and Th1 cell frequencies. Representative dot plots using matching peripheral blood and bone marrow samples from B-ALL patients and healthy donors (HD) were shown. b Statistical data for frequencies of Th17 and Th1 cells within CD4 + T population were shown. c Total RNA was extracted from CD4 + T cells isolated from B-ALL patients and HDs and reverse transcribed into cDNA and subsequently determined for IL-17A and IFN-γ mRNA expression using quantitative PCR. d The frequencies of Th17 cells were significantly decreased in BM when B-ALL patients achieved complete remission (CR). e CD4 + T cells were cultured with or without Nalm-6 cells for 14 days in the presence of OKT3 plus IL-2 (300 units/ml). Then, frequencies of Th17 cells were determined after stimulation with PMA plus ionomycin
    Figure Legend Snippet: Th17 cells increase with reduced Th1 cells in freshly isolated PBMCs and BMMCs from patients with B-ALL. a PBMCs and BMMCs were separated and stimulated with PMA and ionomycin for 5 h in the presence of brefeldin A and subsequently stained with antibodies against CD3, CD8, intracellular IL-17A or IFN-γ. Flow cytometric analysis was used to determine the Th17 and Th1 cell frequencies. Representative dot plots using matching peripheral blood and bone marrow samples from B-ALL patients and healthy donors (HD) were shown. b Statistical data for frequencies of Th17 and Th1 cells within CD4 + T population were shown. c Total RNA was extracted from CD4 + T cells isolated from B-ALL patients and HDs and reverse transcribed into cDNA and subsequently determined for IL-17A and IFN-γ mRNA expression using quantitative PCR. d The frequencies of Th17 cells were significantly decreased in BM when B-ALL patients achieved complete remission (CR). e CD4 + T cells were cultured with or without Nalm-6 cells for 14 days in the presence of OKT3 plus IL-2 (300 units/ml). Then, frequencies of Th17 cells were determined after stimulation with PMA plus ionomycin

    Techniques Used: Isolation, Staining, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction, Cell Culture

    2) Product Images from "Synthetic Peptides Elicit Strong Cellular Immunity in Visceral Leishmaniasis Natural Reservoir and Contribute to Long-Lasting Polyfunctional T-Cells in BALB/c Mice"

    Article Title: Synthetic Peptides Elicit Strong Cellular Immunity in Visceral Leishmaniasis Natural Reservoir and Contribute to Long-Lasting Polyfunctional T-Cells in BALB/c Mice

    Journal: Vaccines

    doi: 10.3390/vaccines7040162

    In vitro proliferation and IFN-γ production by T-lymphocytes in peripheral blood mononuclear cells (PBMC) of L. infantum naturally infected dogs after stimuli with predicted peptides. The spots represent indexes of T-cell responses to Leishmania infantum peptide pools in PBMC of naturally infected and asymptomatic dogs ( n = 5 in gray circles). Proliferation index of CD4 + ( a ) and CD8 + ( b ) lymphocytes after peptide pool stimuli. ( c ) Index of IFN-γ producing by CD4 + and CD8 + ( d ) lymphocytes after peptide pool stimuli. Indexes were calculated based on SLA (soluble Leishmania antigen)-stimulated (SC) cultures divided by the control culture (CC). Cut-off (dashed line) for responses was calculated by the control culture indexes (cut-off = 1).
    Figure Legend Snippet: In vitro proliferation and IFN-γ production by T-lymphocytes in peripheral blood mononuclear cells (PBMC) of L. infantum naturally infected dogs after stimuli with predicted peptides. The spots represent indexes of T-cell responses to Leishmania infantum peptide pools in PBMC of naturally infected and asymptomatic dogs ( n = 5 in gray circles). Proliferation index of CD4 + ( a ) and CD8 + ( b ) lymphocytes after peptide pool stimuli. ( c ) Index of IFN-γ producing by CD4 + and CD8 + ( d ) lymphocytes after peptide pool stimuli. Indexes were calculated based on SLA (soluble Leishmania antigen)-stimulated (SC) cultures divided by the control culture (CC). Cut-off (dashed line) for responses was calculated by the control culture indexes (cut-off = 1).

    Techniques Used: In Vitro, Infection

    3) Product Images from "Potent In Vitro Activity of Citrus aurantium Essential Oil and Vitis vinifera Hydrolate Against Gut Yeast Isolates from Irritable Bowel Syndrome Patients—The Right Mix for Potential Therapeutic Use"

    Article Title: Potent In Vitro Activity of Citrus aurantium Essential Oil and Vitis vinifera Hydrolate Against Gut Yeast Isolates from Irritable Bowel Syndrome Patients—The Right Mix for Potential Therapeutic Use

    Journal: Nutrients

    doi: 10.3390/nu12051329

    Variation of the amount of cytokines IL-10 and TNF-α in the medium culture of PBMCs treated with scalar dilutions of the V. vinifera cv Italia Hy alone or in combination with EO and/or LPS. The circle-shaped indicators refer to the samples treated with the Hy only; the squares are referred to the samples treated with the Hy plus LPS, while the triangles show the samples treated with Hy plus the EO of C. aurantium var. amara plus LPS. The white rhombus is the untreated control and the black rhombus is the control treated with LPS. Letters a , b , c and d indicate samples treated with scalar dilution of the Hy (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively). Letters e , f , g and h are referred to samples treated with scalar dilution of the Hy (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively) and LPS (1 μgr/mL). Letters i , l , m and n are referred to PBMCs treated with a 1:100 v / v mixture of the Hy, (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively) and the EO (0.50% v / v , 0.25% v / v , 0.12% v / v and 0.06% v / v respectively) plus LPS.
    Figure Legend Snippet: Variation of the amount of cytokines IL-10 and TNF-α in the medium culture of PBMCs treated with scalar dilutions of the V. vinifera cv Italia Hy alone or in combination with EO and/or LPS. The circle-shaped indicators refer to the samples treated with the Hy only; the squares are referred to the samples treated with the Hy plus LPS, while the triangles show the samples treated with Hy plus the EO of C. aurantium var. amara plus LPS. The white rhombus is the untreated control and the black rhombus is the control treated with LPS. Letters a , b , c and d indicate samples treated with scalar dilution of the Hy (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively). Letters e , f , g and h are referred to samples treated with scalar dilution of the Hy (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively) and LPS (1 μgr/mL). Letters i , l , m and n are referred to PBMCs treated with a 1:100 v / v mixture of the Hy, (50% v / v , 25% v / v , 12.50% v / v and 0.06% v / v respectively) and the EO (0.50% v / v , 0.25% v / v , 0.12% v / v and 0.06% v / v respectively) plus LPS.

    Techniques Used:

    Related Articles

    Lysis:

    Article Title: Preliminary Trichinella spiralis Infection Ameliorates Subsequent RSV Infection-Induced Inflammatory Response
    Article Snippet: Inflammatory cell influx in the bronchoalveolar lavage fluid (BALF) were quantified under the microscope using a hemocytometer. .. Collected BALF samples from the murine lungs were centrifuged at 1200 RPM, 4 °C, 3 min, followed by red blood cell lysis using RBC lysis buffer (Sigma Aldrich, St. Louis, MO, USA). .. After centrifugation, supernatants were aspirated and cells were resuspended in 100 uL PBS, then counted.

    Article Title: Peptides released from reovirus outer capsid form membrane pores that recruit virus particles
    Article Snippet: .. Times of preconversion for ISVPs, dpSVPs, and WTprcs were measured by incubating RBC+ reactions at 37°C and checking for RBC lysis every 30–45 s. Time of preconversion for N42Aprcs was measured by taking aliquots of RBC+ reactions every minute for several minutes to tubes with trypsin (Sigma-Aldrich) (final concentration 200 μg/ml in VB), incubating on ice for 30–40 min, and analysing by SDS–PAGE and Coomassie Blue staining. ..

    Article Title: Activating Transcription Factor 4 Is Critical for Proliferation and Survival in Primary Bone Marrow Stromal Cells and Calvarial Osteoblasts
    Article Snippet: We first determined the effects of ATF4 deficiency on the numbers of total nucleated bone marrow cells, bone marrow from 6-week-old male wt and Atf4 −/− mice (6/group) were isolated. .. After lysing the red blood cells (RBCs) using the RBC lysis buffer (Sigma), the remaining nucleated bone marrow cells were directly counted using a hemacytometer. .. As shown in , although the long bones of Atf4 −/− mice were significantly shorter and thinner than those of wt mice, surprisingly, we found no significant difference in the numbers of total nucleated bone marrow cells between wt and Atf4 −/− mice.

    Isolation:

    Article Title: Induction of Noonan syndrome-specific human-induced pluripotent stem cells under serum-, feeder-, and integration-free conditions
    Article Snippet: Both WT-DPC-hiPSCs and NS-DPC-hiPSCs were infected following the same protocol. .. Isolation of peripheral blood mononuclear cells (PBMCs) and infection protocol of SeVdp under serum-free conditions Induction of PBMC-hiPSCs was performed as reported previously (Hamada et al .). .. Briefly, PBMCs were prepared by density gradient centrifugation in a Histopaque 1077 (Sigma-Aldrich) and were cultured in RD6F serum-free medium supplemented with IL-2 (CELEUK, Takeda Pharm., Osaka, Japan) for 6 d at 37°C in a humidified atmosphere of 95% air/5% CO2 .

    Infection:

    Article Title: Induction of Noonan syndrome-specific human-induced pluripotent stem cells under serum-, feeder-, and integration-free conditions
    Article Snippet: Both WT-DPC-hiPSCs and NS-DPC-hiPSCs were infected following the same protocol. .. Isolation of peripheral blood mononuclear cells (PBMCs) and infection protocol of SeVdp under serum-free conditions Induction of PBMC-hiPSCs was performed as reported previously (Hamada et al .). .. Briefly, PBMCs were prepared by density gradient centrifugation in a Histopaque 1077 (Sigma-Aldrich) and were cultured in RD6F serum-free medium supplemented with IL-2 (CELEUK, Takeda Pharm., Osaka, Japan) for 6 d at 37°C in a humidified atmosphere of 95% air/5% CO2 .

    Concentration Assay:

    Article Title: Peptides released from reovirus outer capsid form membrane pores that recruit virus particles
    Article Snippet: .. Times of preconversion for ISVPs, dpSVPs, and WTprcs were measured by incubating RBC+ reactions at 37°C and checking for RBC lysis every 30–45 s. Time of preconversion for N42Aprcs was measured by taking aliquots of RBC+ reactions every minute for several minutes to tubes with trypsin (Sigma-Aldrich) (final concentration 200 μg/ml in VB), incubating on ice for 30–40 min, and analysing by SDS–PAGE and Coomassie Blue staining. ..

    SDS Page:

    Article Title: Peptides released from reovirus outer capsid form membrane pores that recruit virus particles
    Article Snippet: .. Times of preconversion for ISVPs, dpSVPs, and WTprcs were measured by incubating RBC+ reactions at 37°C and checking for RBC lysis every 30–45 s. Time of preconversion for N42Aprcs was measured by taking aliquots of RBC+ reactions every minute for several minutes to tubes with trypsin (Sigma-Aldrich) (final concentration 200 μg/ml in VB), incubating on ice for 30–40 min, and analysing by SDS–PAGE and Coomassie Blue staining. ..

    Staining:

    Article Title: Peptides released from reovirus outer capsid form membrane pores that recruit virus particles
    Article Snippet: .. Times of preconversion for ISVPs, dpSVPs, and WTprcs were measured by incubating RBC+ reactions at 37°C and checking for RBC lysis every 30–45 s. Time of preconversion for N42Aprcs was measured by taking aliquots of RBC+ reactions every minute for several minutes to tubes with trypsin (Sigma-Aldrich) (final concentration 200 μg/ml in VB), incubating on ice for 30–40 min, and analysing by SDS–PAGE and Coomassie Blue staining. ..

    Article Title: An accurate, precise method for general labeling of extracellular vesicles
    Article Snippet: .. Validation Approach 2 – Permeabilization and general membrane staining To demonstrate that calcein AM labels intact EVs-but not membrane fragments or other debris-we permeabilized the plasma, RBC, and HAEC EVs with two different agents: Triton X-100 (Sigma-Aldrich, cat. T-8787) and saponin (Sigma-Aldrich, cat. 47036-50G-F). ..

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  • 98
    Millipore pbmcs proliferation
    <t>MSCs</t> from pericardial and subcutaneous adipose tissue equally suppress T‐cell proliferation. (A) : Representative example of a flow cytometry proliferation analysis of monocyte depleted peripheral blood mononuclear cells in coculture with subcutaneous or pericardial MSCs. MSCs from subcutaneous and pericardial fat have similar ability to suppress activated T‐cells' proliferation (B) and to support T‐cell viability (C) ( n = 8). Abbreviations: 7 AAD, 7‐aminoactinomycin D; adMSCs, adipose tissue‐derived MSCs; EI, expansion index; CFSE, carboxyfluorescein succinimidyl ester; FSC‐A: forward scatter area; SSC‐A: side scatter area; SSC‐H: side scatter height; SSC‐W: side scatter width.
    Pbmcs Proliferation, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs proliferation/product/Millipore
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pbmcs proliferation - by Bioz Stars, 2021-03
    98/100 stars
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    99
    Millipore peripheral blood mononuclear cells
    Raman fingerprints of extracellular vesicles. Raman spectra of trophoblast (a) and <t>peripheral</t> <t>mononuclear</t> <t>blood</t> cell (PBMC) (b) extracellular vesicles, n = 30–35 . The spectral shift positions are highlighted in cyan color and indicated by red numbers.
    Peripheral Blood Mononuclear Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    peripheral blood mononuclear cells - by Bioz Stars, 2021-03
    99/100 stars
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    Image Search Results


    MSCs from pericardial and subcutaneous adipose tissue equally suppress T‐cell proliferation. (A) : Representative example of a flow cytometry proliferation analysis of monocyte depleted peripheral blood mononuclear cells in coculture with subcutaneous or pericardial MSCs. MSCs from subcutaneous and pericardial fat have similar ability to suppress activated T‐cells' proliferation (B) and to support T‐cell viability (C) ( n = 8). Abbreviations: 7 AAD, 7‐aminoactinomycin D; adMSCs, adipose tissue‐derived MSCs; EI, expansion index; CFSE, carboxyfluorescein succinimidyl ester; FSC‐A: forward scatter area; SSC‐A: side scatter area; SSC‐H: side scatter height; SSC‐W: side scatter width.

    Journal: Stem Cells Translational Medicine

    Article Title: A Proinflammatory Secretome Mediates the Impaired Immunopotency of Human Mesenchymal Stromal Cells in Elderly Patients with Atherosclerosis

    doi: 10.1002/sctm.16-0221

    Figure Lengend Snippet: MSCs from pericardial and subcutaneous adipose tissue equally suppress T‐cell proliferation. (A) : Representative example of a flow cytometry proliferation analysis of monocyte depleted peripheral blood mononuclear cells in coculture with subcutaneous or pericardial MSCs. MSCs from subcutaneous and pericardial fat have similar ability to suppress activated T‐cells' proliferation (B) and to support T‐cell viability (C) ( n = 8). Abbreviations: 7 AAD, 7‐aminoactinomycin D; adMSCs, adipose tissue‐derived MSCs; EI, expansion index; CFSE, carboxyfluorescein succinimidyl ester; FSC‐A: forward scatter area; SSC‐A: side scatter area; SSC‐H: side scatter height; SSC‐W: side scatter width.

    Article Snippet: To assess the effect of MSCs on suppressing monocyte‐depleted PBMCs proliferation, PBMCs were labeled with 10 uM carboxyfluorescein succinimidyl ester (CFSE) (Sigma), stimulated with anti‐CD3/CD28 beads (1 bead per cell) (Dynabeads Human T‐Activator CD3/CD28, Life Technologies) and cultured for 4 days with MSCs.

    Techniques: Flow Cytometry, Cytometry, Derivative Assay

    Raman fingerprints of extracellular vesicles. Raman spectra of trophoblast (a) and peripheral mononuclear blood cell (PBMC) (b) extracellular vesicles, n = 30–35 . The spectral shift positions are highlighted in cyan color and indicated by red numbers.

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: Raman fingerprints of extracellular vesicles. Raman spectra of trophoblast (a) and peripheral mononuclear blood cell (PBMC) (b) extracellular vesicles, n = 30–35 . The spectral shift positions are highlighted in cyan color and indicated by red numbers.

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques:

    PCA and LDA analyses of the extracellular vesicles derived from peripheral mononuclear blood cells (P001, P002 and P003) and trophoblast cells (T001, T002 and T003). Scatter plot of the PCA results show the PC1 and PC2 scores assigned to each spectrum ( N = 90 ) (a). Linear discriminant analysis, the first 10 PC loadings calculated by means of PCA were used for LDA. Each spot represents a single spectrum. The crosses indicate the mean canonical observation score obtained for each group ( N = 90 ) (b). 2D PCA (c) and LDA (d) plots of PBMC-derived vesicles from three different cows, and 2D PCA (e) and LDA (f) plots of trophoblast-derived vesicles from three different animal individuals.

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: PCA and LDA analyses of the extracellular vesicles derived from peripheral mononuclear blood cells (P001, P002 and P003) and trophoblast cells (T001, T002 and T003). Scatter plot of the PCA results show the PC1 and PC2 scores assigned to each spectrum ( N = 90 ) (a). Linear discriminant analysis, the first 10 PC loadings calculated by means of PCA were used for LDA. Each spot represents a single spectrum. The crosses indicate the mean canonical observation score obtained for each group ( N = 90 ) (b). 2D PCA (c) and LDA (d) plots of PBMC-derived vesicles from three different cows, and 2D PCA (e) and LDA (f) plots of trophoblast-derived vesicles from three different animal individuals.

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques: Derivative Assay

    Principal component analysis (PCA) loading plots corresponding to PC1, PC2, and PC3 from the data of Fig 6A for extracellular vesicles derived from peripheral mononuclear blood cells and trophoblast cells.

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: Principal component analysis (PCA) loading plots corresponding to PC1, PC2, and PC3 from the data of Fig 6A for extracellular vesicles derived from peripheral mononuclear blood cells and trophoblast cells.

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques: Derivative Assay

    Raman fingerprint of extracellular vesicles (EVs) isolated from peripheral mononuclear blood cells (PBMC) and trophoblast cells. The solid line indicates the average of 90 spectra ± 1 standard deviation (shaded grey areas). The areas highlighted in cyan color showed the intensity differences. The peaks marked in red only exist in PBMC-derived EVs, while the peaks in blue only exist in trophoblast-derived EVs (a). Mean Raman peak intensity analysis of PBMC and trophoblast-derived EVs, N = 90 , * means P

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: Raman fingerprint of extracellular vesicles (EVs) isolated from peripheral mononuclear blood cells (PBMC) and trophoblast cells. The solid line indicates the average of 90 spectra ± 1 standard deviation (shaded grey areas). The areas highlighted in cyan color showed the intensity differences. The peaks marked in red only exist in PBMC-derived EVs, while the peaks in blue only exist in trophoblast-derived EVs (a). Mean Raman peak intensity analysis of PBMC and trophoblast-derived EVs, N = 90 , * means P

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques: Isolation, Standard Deviation, Derivative Assay

    Generation, isolation, and characterization of epidermal growth factor receptor (EGFR)‐specific chimeric antigen receptor (CAR)‐engineered natural killer (NK) cells (EGFR‐CAR NK cells). (A) Flow cytometric analysis of phenotypic and subset composition of peripheral blood mononuclear cells (PBMCs) labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE. (B‐C) Flow cytometric analysis of phenotypic and subset composition of NK cells labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE, and anti‐CD69‐APC‐Cy7. (D) The percentage of CD3‐/CD56 + cells in day 0 PBMCs and day14 PBMC culture. (E) Real‐time PCR and (F) Western blotting analyses of the expression of exogenous CD3ζ in the non‐transduced NK cells, Con‐CAR NK cells, EGFR‐CAR‐1 NK cells, and EGFR‐CAR‐2 NK cells. β‐actin was used as an endogenous control. (G) The transduced NK cells stained with IgG‐FITC and EGFR‐FITC antibodies were detected by flow cytometry

    Journal: Cell Proliferation

    Article Title: Targeting epidermal growth factor‐overexpressing triple‐negative breast cancer by natural killer cells expressing a specific chimeric antigen receptor, et al. Targeting epidermal growth factor‐overexpressing triple‐negative breast cancer by natural killer cells expressing a specific chimeric antigen receptor

    doi: 10.1111/cpr.12858

    Figure Lengend Snippet: Generation, isolation, and characterization of epidermal growth factor receptor (EGFR)‐specific chimeric antigen receptor (CAR)‐engineered natural killer (NK) cells (EGFR‐CAR NK cells). (A) Flow cytometric analysis of phenotypic and subset composition of peripheral blood mononuclear cells (PBMCs) labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE. (B‐C) Flow cytometric analysis of phenotypic and subset composition of NK cells labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE, and anti‐CD69‐APC‐Cy7. (D) The percentage of CD3‐/CD56 + cells in day 0 PBMCs and day14 PBMC culture. (E) Real‐time PCR and (F) Western blotting analyses of the expression of exogenous CD3ζ in the non‐transduced NK cells, Con‐CAR NK cells, EGFR‐CAR‐1 NK cells, and EGFR‐CAR‐2 NK cells. β‐actin was used as an endogenous control. (G) The transduced NK cells stained with IgG‐FITC and EGFR‐FITC antibodies were detected by flow cytometry

    Article Snippet: Peripheral blood mononuclear cells were isolated from the whole blood of healthy donors by Ficoll density gradient centrifugation.

    Techniques: Isolation, Labeling, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Staining, Flow Cytometry

    Human and murine ILC2 gating strategy. A. Human ILC2s were isolated from peripheral blood of healthy donors PBMCs or umbilical cord blood samples CBMCs and stained with antibodies against CD45 and lineage markers. Human ILC2s were sorted by the BD FACSAria cell sorter as CD45 + Lin - CRTH2 + CD127 + cells. B. Murine ILC2s were isolated from mouse lungs treated with recombinant IL-33 protein (250ng/mouse, i.t. ) and stained with antibodies against CD45 and lineage markers as described in the Materials and Methods. Murine ILC2s were sorted by the BD FACSAria cell sorter as CD45 + Lin - T1/ST2 + cells. The purity of sorted ILC2s was determined to be greater than 95%.

    Journal: bioRxiv

    Article Title: Non-canonical Activation of Human Group 2 Innate Lymphoid Cells by TLR4 Signaling

    doi: 10.1101/2020.10.29.361345

    Figure Lengend Snippet: Human and murine ILC2 gating strategy. A. Human ILC2s were isolated from peripheral blood of healthy donors PBMCs or umbilical cord blood samples CBMCs and stained with antibodies against CD45 and lineage markers. Human ILC2s were sorted by the BD FACSAria cell sorter as CD45 + Lin - CRTH2 + CD127 + cells. B. Murine ILC2s were isolated from mouse lungs treated with recombinant IL-33 protein (250ng/mouse, i.t. ) and stained with antibodies against CD45 and lineage markers as described in the Materials and Methods. Murine ILC2s were sorted by the BD FACSAria cell sorter as CD45 + Lin - T1/ST2 + cells. The purity of sorted ILC2s was determined to be greater than 95%.

    Article Snippet: Peripheral or Cord Blood Mononuclear Cells (PBMCs or CBMCs) were isolated from diluted umbilical cord blood (1:2) by density gradient centrifugation using density gradient medium, Histopaque® (Sigma Aldrich) and SepMateTM 50 mL tubes (STEMCELL Technologies) ( ).

    Techniques: Isolation, Staining, Recombinant