rpmi 1640 medium Search Results


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  • 93
    Lonza rpmi 1640
    Rpmi 1640, supplied by Lonza, used in various techniques. Bioz Stars score: 93/100, based on 5862 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC rpmi 1640 medium
    Rpmi 1640 Medium, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 2163 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rpmi 1640 medium
    Rpmi 1640 Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 100450 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rpmi 1640
    Rpmi 1640, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 91082 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore rpmi 1640 medium
    Rpmi 1640 Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 30869 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rpmi medium
    Rpmi Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 21095 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare rpmi 1640 medium
    Rpmi 1640 Medium, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 12494 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rpmi1640
    Rpmi1640, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 7158 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Lonza rpmi 1640 medium
    Rpmi 1640 Medium, supplied by Lonza, used in various techniques. Bioz Stars score: 94/100, based on 5133 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Mediatech rpmi 1640 medium
    Figure 6.  Cell cycle kinetics of cancer cells during seeding on Gelffoam®. FUCCI-expressing cancer cells (5 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) Low-magnification image of an overview
    Rpmi 1640 Medium, supplied by Mediatech, used in various techniques. Bioz Stars score: 94/100, based on 2810 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Corning Life Sciences rpmi 1640 medium
    MTR knockout reduces SAM levels but not methionine levels. (A) Proliferation rates of A549 cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM methionine to supplement the 100 μM methionine present in <t>RPMI-1640</t> culture media. (B) LC/MS measurement of intracellular methionine, lysine, and serine levels in A549 cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. Data are normalized to the total protein content of cells in each condition and to an internal standard. (C) LC/MS measurement of intracellular SAM and SAH levels in A549 cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. Data are normalized to the total protein content of cells in each condition and to an internal standard. (D) Western blots to assess levels of mono- or dimethyl lysine-containing proteins, RPS6 phosphorylated at S235/6, total RPS6, S6K phosphorylated at T389, and total S6K in A549 cells without (+EV) or with MTR expression (+MTR). Phosphorylation index is defined as the phosphorylated protein antibody signal divided by the same total protein antibody signal in each lane. (E) Proliferation rates of A549 cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM SAM. Mean +/- SD is displayed for all panels. p values indicated on all panels are derived from two-tailed, unpaired Welch’s t tests.
    Rpmi 1640 Medium, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 94/100, based on 1992 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cellgro rpmi 1640 medium
    MTR knockout reduces SAM levels but not methionine levels. (A) Proliferation rates of A549 cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM methionine to supplement the 100 μM methionine present in <t>RPMI-1640</t> culture media. (B) LC/MS measurement of intracellular methionine, lysine, and serine levels in A549 cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. Data are normalized to the total protein content of cells in each condition and to an internal standard. (C) LC/MS measurement of intracellular SAM and SAH levels in A549 cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. Data are normalized to the total protein content of cells in each condition and to an internal standard. (D) Western blots to assess levels of mono- or dimethyl lysine-containing proteins, RPS6 phosphorylated at S235/6, total RPS6, S6K phosphorylated at T389, and total S6K in A549 cells without (+EV) or with MTR expression (+MTR). Phosphorylation index is defined as the phosphorylated protein antibody signal divided by the same total protein antibody signal in each lane. (E) Proliferation rates of A549 cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM SAM. Mean +/- SD is displayed for all panels. p values indicated on all panels are derived from two-tailed, unpaired Welch’s t tests.
    Rpmi 1640 Medium, supplied by Cellgro, used in various techniques. Bioz Stars score: 94/100, based on 1932 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biochrom rpmi 1640 medium
    Viability of intracellular Leishmania in PMN. PMN were coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 3 h at 37°C in <t>RPMI</t> 1640 medium supplemented with either 20% fresh autologous serum or 20% heat-inactivated FCS (hi-FCS) or without serum supplementation. PMN were then separated from nonphagocytosed L. major promastigotes by fluorescence-activated cell sorting and subsequently incubated for 24 h at 37°C in complete RPMI 1640 medium containing 10% FCS. The viability of L. major in PMN was then assessed by a limiting-dilution Leishmania culture assay. The y axis shows the mean and standard deviation of the three parallel dilutions of PMN.
    Rpmi 1640 Medium, supplied by Biochrom, used in various techniques. Bioz Stars score: 93/100, based on 1760 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher phenol red
    Viability of intracellular Leishmania in PMN. PMN were coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 3 h at 37°C in <t>RPMI</t> 1640 medium supplemented with either 20% fresh autologous serum or 20% heat-inactivated FCS (hi-FCS) or without serum supplementation. PMN were then separated from nonphagocytosed L. major promastigotes by fluorescence-activated cell sorting and subsequently incubated for 24 h at 37°C in complete RPMI 1640 medium containing 10% FCS. The viability of L. major in PMN was then assessed by a limiting-dilution Leishmania culture assay. The y axis shows the mean and standard deviation of the three parallel dilutions of PMN.
    Phenol Red, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 12676 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Figure 6.  Cell cycle kinetics of cancer cells during seeding on Gelffoam®. FUCCI-expressing cancer cells (5 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) Low-magnification image of an overview

    Journal: Cell Cycle

    Article Title: Invading cancer cells are predominantly in G0/G1 resulting in chemoresistance demonstrated by real-time FUCCI imaging

    doi: 10.4161/cc.27818

    Figure Lengend Snippet: Figure 6. Cell cycle kinetics of cancer cells during seeding on Gelffoam®. FUCCI-expressing cancer cells (5 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) Low-magnification image of an overview

    Article Snippet: FUCCI-expressing MKN45 cells were seeded on plastic plates for 2-dimensional culture in RPMI-1640 medium (Mediatech).

    Techniques: Expressing

    Figure 5.  Comparison of migration velocity of cancer cells in G 0 /G 1  phase and S/G 2 /M phases during cell division. FUCCI-expressing cancer cells (5 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) Cancer

    Journal: Cell Cycle

    Article Title: Invading cancer cells are predominantly in G0/G1 resulting in chemoresistance demonstrated by real-time FUCCI imaging

    doi: 10.4161/cc.27818

    Figure Lengend Snippet: Figure 5. Comparison of migration velocity of cancer cells in G 0 /G 1 phase and S/G 2 /M phases during cell division. FUCCI-expressing cancer cells (5 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) Cancer

    Article Snippet: FUCCI-expressing MKN45 cells were seeded on plastic plates for 2-dimensional culture in RPMI-1640 medium (Mediatech).

    Techniques: Migration, Expressing

    Figure 7.  Chemotherapy does not kill or inhibit the movement of single invading cancer cells in G 0 /G 1  phase. FUCCI-expressing cancer cells (2 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. Three days after

    Journal: Cell Cycle

    Article Title: Invading cancer cells are predominantly in G0/G1 resulting in chemoresistance demonstrated by real-time FUCCI imaging

    doi: 10.4161/cc.27818

    Figure Lengend Snippet: Figure 7. Chemotherapy does not kill or inhibit the movement of single invading cancer cells in G 0 /G 1 phase. FUCCI-expressing cancer cells (2 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. Three days after

    Article Snippet: FUCCI-expressing MKN45 cells were seeded on plastic plates for 2-dimensional culture in RPMI-1640 medium (Mediatech).

    Techniques: Expressing

    Figure 4.  Behavior of individual FUCCI-expressing cancer cells cultured on Gelfoam®. FUCCI-expressing cancer cells (5 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) Low-magnification image of

    Journal: Cell Cycle

    Article Title: Invading cancer cells are predominantly in G0/G1 resulting in chemoresistance demonstrated by real-time FUCCI imaging

    doi: 10.4161/cc.27818

    Figure Lengend Snippet: Figure 4. Behavior of individual FUCCI-expressing cancer cells cultured on Gelfoam®. FUCCI-expressing cancer cells (5 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) Low-magnification image of

    Article Snippet: FUCCI-expressing MKN45 cells were seeded on plastic plates for 2-dimensional culture in RPMI-1640 medium (Mediatech).

    Techniques: Expressing, Cell Culture

    Figure 2.  Cell cycle distribution of invasive and noninvasive cancer cells. FUCCI-expressing cancer cells (1 × 10 7 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) High-magnification real-time images of a small

    Journal: Cell Cycle

    Article Title: Invading cancer cells are predominantly in G0/G1 resulting in chemoresistance demonstrated by real-time FUCCI imaging

    doi: 10.4161/cc.27818

    Figure Lengend Snippet: Figure 2. Cell cycle distribution of invasive and noninvasive cancer cells. FUCCI-expressing cancer cells (1 × 10 7 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) High-magnification real-time images of a small

    Article Snippet: FUCCI-expressing MKN45 cells were seeded on plastic plates for 2-dimensional culture in RPMI-1640 medium (Mediatech).

    Techniques: Expressing

    Figure 3.  Invasive cancer cells are predominantly in G 0 /G 1 . FUCCI-expressing cancer cells (5 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) High-magnification real-time images of invading cancer cells

    Journal: Cell Cycle

    Article Title: Invading cancer cells are predominantly in G0/G1 resulting in chemoresistance demonstrated by real-time FUCCI imaging

    doi: 10.4161/cc.27818

    Figure Lengend Snippet: Figure 3. Invasive cancer cells are predominantly in G 0 /G 1 . FUCCI-expressing cancer cells (5 × 10 6 ) were placed on Gelfoam® (1 × 1 cm) in RPMI 1640 medium. ( A ) High-magnification real-time images of invading cancer cells

    Article Snippet: FUCCI-expressing MKN45 cells were seeded on plastic plates for 2-dimensional culture in RPMI-1640 medium (Mediatech).

    Techniques: Expressing

    MTR knockout reduces SAM levels but not methionine levels. (A) Proliferation rates of A549 cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM methionine to supplement the 100 μM methionine present in RPMI-1640 culture media. (B) LC/MS measurement of intracellular methionine, lysine, and serine levels in A549 cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. Data are normalized to the total protein content of cells in each condition and to an internal standard. (C) LC/MS measurement of intracellular SAM and SAH levels in A549 cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. Data are normalized to the total protein content of cells in each condition and to an internal standard. (D) Western blots to assess levels of mono- or dimethyl lysine-containing proteins, RPS6 phosphorylated at S235/6, total RPS6, S6K phosphorylated at T389, and total S6K in A549 cells without (+EV) or with MTR expression (+MTR). Phosphorylation index is defined as the phosphorylated protein antibody signal divided by the same total protein antibody signal in each lane. (E) Proliferation rates of A549 cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM SAM. Mean +/- SD is displayed for all panels. p values indicated on all panels are derived from two-tailed, unpaired Welch’s t tests.

    Journal: bioRxiv

    Article Title: Methionine synthase is essential for cancer cell proliferation in physiological folate environments

    doi: 10.1101/2020.06.12.149005

    Figure Lengend Snippet: MTR knockout reduces SAM levels but not methionine levels. (A) Proliferation rates of A549 cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM methionine to supplement the 100 μM methionine present in RPMI-1640 culture media. (B) LC/MS measurement of intracellular methionine, lysine, and serine levels in A549 cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. Data are normalized to the total protein content of cells in each condition and to an internal standard. (C) LC/MS measurement of intracellular SAM and SAH levels in A549 cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. Data are normalized to the total protein content of cells in each condition and to an internal standard. (D) Western blots to assess levels of mono- or dimethyl lysine-containing proteins, RPS6 phosphorylated at S235/6, total RPS6, S6K phosphorylated at T389, and total S6K in A549 cells without (+EV) or with MTR expression (+MTR). Phosphorylation index is defined as the phosphorylated protein antibody signal divided by the same total protein antibody signal in each lane. (E) Proliferation rates of A549 cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM SAM. Mean +/- SD is displayed for all panels. p values indicated on all panels are derived from two-tailed, unpaired Welch’s t tests.

    Article Snippet: Cells were trypsinized, counted and plated into six well dishes (Corning Life Sciences) in 2 mL of RPMI-1640 medium lacking folic acid and incubated overnight.

    Techniques: Knock-Out, Expressing, Cell Culture, Liquid Chromatography with Mass Spectroscopy, Western Blot, Derivative Assay, Two Tailed Test

    MTR expression affects SAM but not methionine metabolism in T.T cells. (A) Proliferation rates of T.T cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM methionine to supplement the 100 μM methionine present in RPMI-1640 culture media. (B) LC/MS measurement of intracellular methionine, lysine, and serine levels in T.T cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. (C) LC/MS measurement of intracellular SAM and SAH levels in T.T cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. (D) Western blots to assess levels of mono- or dimethyl lysine-containing proteins, RPS6 phosphorylated at S235/6, total RPS6, S6K phosphorylated at T389, and total S6K in T.T cells without (+EV) or with MTR expression (+MTR). Phosphorylation index is defined as the phosphorylated protein antibody signal divided by the same total protein antibody signal in each lane. (E) Proliferation rates of T.T cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM SAM. Mean +/- SD is displayed for all panels. p values indicated on all panels are derived from two-tailed, unpaired Welch’s t tests. For LC/MS measurements, data are normalized to the total protein content of cells in each condition and to an internal standard.

    Journal: bioRxiv

    Article Title: Methionine synthase is essential for cancer cell proliferation in physiological folate environments

    doi: 10.1101/2020.06.12.149005

    Figure Lengend Snippet: MTR expression affects SAM but not methionine metabolism in T.T cells. (A) Proliferation rates of T.T cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM methionine to supplement the 100 μM methionine present in RPMI-1640 culture media. (B) LC/MS measurement of intracellular methionine, lysine, and serine levels in T.T cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. (C) LC/MS measurement of intracellular SAM and SAH levels in T.T cells without (+EV) or with MTR expression (+MTR) cultured for 4 days in the indicated folate. (D) Western blots to assess levels of mono- or dimethyl lysine-containing proteins, RPS6 phosphorylated at S235/6, total RPS6, S6K phosphorylated at T389, and total S6K in T.T cells without (+EV) or with MTR expression (+MTR). Phosphorylation index is defined as the phosphorylated protein antibody signal divided by the same total protein antibody signal in each lane. (E) Proliferation rates of T.T cells without (+EV) or with MTR expression (+MTR) cultured in the indicated folate with or without the addition of 1 mM SAM. Mean +/- SD is displayed for all panels. p values indicated on all panels are derived from two-tailed, unpaired Welch’s t tests. For LC/MS measurements, data are normalized to the total protein content of cells in each condition and to an internal standard.

    Article Snippet: Cells were trypsinized, counted and plated into six well dishes (Corning Life Sciences) in 2 mL of RPMI-1640 medium lacking folic acid and incubated overnight.

    Techniques: Expressing, Cell Culture, Liquid Chromatography with Mass Spectroscopy, Western Blot, Derivative Assay, Two Tailed Test

    Viability of intracellular Leishmania in PMN. PMN were coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 3 h at 37°C in RPMI 1640 medium supplemented with either 20% fresh autologous serum or 20% heat-inactivated FCS (hi-FCS) or without serum supplementation. PMN were then separated from nonphagocytosed L. major promastigotes by fluorescence-activated cell sorting and subsequently incubated for 24 h at 37°C in complete RPMI 1640 medium containing 10% FCS. The viability of L. major in PMN was then assessed by a limiting-dilution Leishmania culture assay. The y axis shows the mean and standard deviation of the three parallel dilutions of PMN.

    Journal: Infection and Immunity

    Article Title: Intracellular Survival of Leishmania major in Neutrophil Granulocytes after Uptake in the Absence of Heat-Labile Serum Factors

    doi:

    Figure Lengend Snippet: Viability of intracellular Leishmania in PMN. PMN were coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 3 h at 37°C in RPMI 1640 medium supplemented with either 20% fresh autologous serum or 20% heat-inactivated FCS (hi-FCS) or without serum supplementation. PMN were then separated from nonphagocytosed L. major promastigotes by fluorescence-activated cell sorting and subsequently incubated for 24 h at 37°C in complete RPMI 1640 medium containing 10% FCS. The viability of L. major in PMN was then assessed by a limiting-dilution Leishmania culture assay. The y axis shows the mean and standard deviation of the three parallel dilutions of PMN.

    Article Snippet: The granulocyte-rich layer of Histopaque 1119 was collected and washed twice in RPMI 1640 medium (Seromed-Biochrom, Berlin, Germany), and the cells were further fractionated on a discontinuous Percoll (Pharmacia, Uppsala, Sweden) gradient consisting of layers with densities of 1,105 g/ml (85%), 1,100 g/ml (80%), 1,093 g/ml (75%), 1,087 g/ml (70%), and 1,081 g/ml (65%).

    Techniques: Fluorescence, FACS, Incubation, Standard Deviation

    Effect of L. major on induction of the oxidative burst by PMN. PMN were coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 10 min at 37°C in RPMI 1640 medium supplemented with either 20% fresh autologous serum or 20% heat-inactivated serum (hi-serum) or without serum supplementation. The oxidative burst in PMN was assessed by using the fluorogenic substrate dihydrorhodamine 123 and measured by flow cytometry (FL-1). The percentage of burst positive cells is shown in the upper left corner.

    Journal: Infection and Immunity

    Article Title: Intracellular Survival of Leishmania major in Neutrophil Granulocytes after Uptake in the Absence of Heat-Labile Serum Factors

    doi:

    Figure Lengend Snippet: Effect of L. major on induction of the oxidative burst by PMN. PMN were coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 10 min at 37°C in RPMI 1640 medium supplemented with either 20% fresh autologous serum or 20% heat-inactivated serum (hi-serum) or without serum supplementation. The oxidative burst in PMN was assessed by using the fluorogenic substrate dihydrorhodamine 123 and measured by flow cytometry (FL-1). The percentage of burst positive cells is shown in the upper left corner.

    Article Snippet: The granulocyte-rich layer of Histopaque 1119 was collected and washed twice in RPMI 1640 medium (Seromed-Biochrom, Berlin, Germany), and the cells were further fractionated on a discontinuous Percoll (Pharmacia, Uppsala, Sweden) gradient consisting of layers with densities of 1,105 g/ml (85%), 1,100 g/ml (80%), 1,093 g/ml (75%), 1,087 g/ml (70%), and 1,081 g/ml (65%).

    Techniques: Flow Cytometry, Cytometry

    Effect of anti-CR3 MAb treatment on the uptake of L. major by PMN. PMN were preincubated with anti-CR3 MAb for 20 min at 37°C, and control cultures were incubated with isotype control MAb. PMN were subsequently washed and coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 2 h at 37°C in RPMI 1640 medium containing 20% fresh autologous serum (A) or 20% heat-inactivated FCS (B). The uptake of parasites was assessed in two experiments (Exp I and Exp II) by microscopic evaluation of Giemsa-stained cytocentrifuge preparations. Exp I and Exp II were performed with cells and serum from two different donors.

    Journal: Infection and Immunity

    Article Title: Intracellular Survival of Leishmania major in Neutrophil Granulocytes after Uptake in the Absence of Heat-Labile Serum Factors

    doi:

    Figure Lengend Snippet: Effect of anti-CR3 MAb treatment on the uptake of L. major by PMN. PMN were preincubated with anti-CR3 MAb for 20 min at 37°C, and control cultures were incubated with isotype control MAb. PMN were subsequently washed and coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 2 h at 37°C in RPMI 1640 medium containing 20% fresh autologous serum (A) or 20% heat-inactivated FCS (B). The uptake of parasites was assessed in two experiments (Exp I and Exp II) by microscopic evaluation of Giemsa-stained cytocentrifuge preparations. Exp I and Exp II were performed with cells and serum from two different donors.

    Article Snippet: The granulocyte-rich layer of Histopaque 1119 was collected and washed twice in RPMI 1640 medium (Seromed-Biochrom, Berlin, Germany), and the cells were further fractionated on a discontinuous Percoll (Pharmacia, Uppsala, Sweden) gradient consisting of layers with densities of 1,105 g/ml (85%), 1,100 g/ml (80%), 1,093 g/ml (75%), 1,087 g/ml (70%), and 1,081 g/ml (65%).

    Techniques: Incubation, Staining

    Kinetics of phagocytosis of L. major promastigotes by PMN in vitro. PMN were incubated with L. major promastigotes at 37°C at a PMN-to-parasite ratio of 1:5 in RPMI 1640 medium supplemented with either 20% fresh autologous serum, 20% heat-inactivated autologous serum (hi-serum), or 20% heat-inactivated FCS (hi-FCS) or without serum supplementation. The percentage of PMN harboring at least one intracellular parasite was assessed by microscopic evaluation of Giemsa-stained cytocentrifuge preparations at the given time points. The data shown are from one experiment representative of four experiments performed.

    Journal: Infection and Immunity

    Article Title: Intracellular Survival of Leishmania major in Neutrophil Granulocytes after Uptake in the Absence of Heat-Labile Serum Factors

    doi:

    Figure Lengend Snippet: Kinetics of phagocytosis of L. major promastigotes by PMN in vitro. PMN were incubated with L. major promastigotes at 37°C at a PMN-to-parasite ratio of 1:5 in RPMI 1640 medium supplemented with either 20% fresh autologous serum, 20% heat-inactivated autologous serum (hi-serum), or 20% heat-inactivated FCS (hi-FCS) or without serum supplementation. The percentage of PMN harboring at least one intracellular parasite was assessed by microscopic evaluation of Giemsa-stained cytocentrifuge preparations at the given time points. The data shown are from one experiment representative of four experiments performed.

    Article Snippet: The granulocyte-rich layer of Histopaque 1119 was collected and washed twice in RPMI 1640 medium (Seromed-Biochrom, Berlin, Germany), and the cells were further fractionated on a discontinuous Percoll (Pharmacia, Uppsala, Sweden) gradient consisting of layers with densities of 1,105 g/ml (85%), 1,100 g/ml (80%), 1,093 g/ml (75%), 1,087 g/ml (70%), and 1,081 g/ml (65%).

    Techniques: In Vitro, Incubation, Staining

    Effect of L. major on the cell surface expression of CD62L and CD66b on PMN. (A) PMN were coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 90 min at 37°C in RPMI 1640 medium supplemented with either 20% fresh autologous serum or 20% heat-inactivated FCS (hi-FCS) or without serum supplementation. PMN were then stained with FITC-conjugated MAb to CD66b and PE-labeled MAb to CD62L and analyzed by flow cytometry (A). (B) Mean fluorescence values of CD66b staining.

    Journal: Infection and Immunity

    Article Title: Intracellular Survival of Leishmania major in Neutrophil Granulocytes after Uptake in the Absence of Heat-Labile Serum Factors

    doi:

    Figure Lengend Snippet: Effect of L. major on the cell surface expression of CD62L and CD66b on PMN. (A) PMN were coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 90 min at 37°C in RPMI 1640 medium supplemented with either 20% fresh autologous serum or 20% heat-inactivated FCS (hi-FCS) or without serum supplementation. PMN were then stained with FITC-conjugated MAb to CD66b and PE-labeled MAb to CD62L and analyzed by flow cytometry (A). (B) Mean fluorescence values of CD66b staining.

    Article Snippet: The granulocyte-rich layer of Histopaque 1119 was collected and washed twice in RPMI 1640 medium (Seromed-Biochrom, Berlin, Germany), and the cells were further fractionated on a discontinuous Percoll (Pharmacia, Uppsala, Sweden) gradient consisting of layers with densities of 1,105 g/ml (85%), 1,100 g/ml (80%), 1,093 g/ml (75%), 1,087 g/ml (70%), and 1,081 g/ml (65%).

    Techniques: Expressing, Staining, Labeling, Flow Cytometry, Cytometry, Fluorescence

    Staining of viable intracellular L. major in PMN with ethidium bromide and acridine orange. PMN were coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 3 h at 37°C in RPMI 1640 medium supplemented with 20% heat-inactivated FCS, separated from nonphagocytosed L. major promastigotes, and subsequently incubated for 24 h at 37°C. The viability of L. major in PMN was then assessed by staining with acridine orange to visualize viable cells or parasites (green staining) and ethidium bromide, which stains dead cells. Viable L. major parasites (arrows) are seen in nonapoptotic viable PMN. The morphological appearance of the nuclei and kinetoplasts is typical for intact parasites.

    Journal: Infection and Immunity

    Article Title: Intracellular Survival of Leishmania major in Neutrophil Granulocytes after Uptake in the Absence of Heat-Labile Serum Factors

    doi:

    Figure Lengend Snippet: Staining of viable intracellular L. major in PMN with ethidium bromide and acridine orange. PMN were coincubated with L. major promastigotes at a PMN-to-parasite ratio of 1:5 for 3 h at 37°C in RPMI 1640 medium supplemented with 20% heat-inactivated FCS, separated from nonphagocytosed L. major promastigotes, and subsequently incubated for 24 h at 37°C. The viability of L. major in PMN was then assessed by staining with acridine orange to visualize viable cells or parasites (green staining) and ethidium bromide, which stains dead cells. Viable L. major parasites (arrows) are seen in nonapoptotic viable PMN. The morphological appearance of the nuclei and kinetoplasts is typical for intact parasites.

    Article Snippet: The granulocyte-rich layer of Histopaque 1119 was collected and washed twice in RPMI 1640 medium (Seromed-Biochrom, Berlin, Germany), and the cells were further fractionated on a discontinuous Percoll (Pharmacia, Uppsala, Sweden) gradient consisting of layers with densities of 1,105 g/ml (85%), 1,100 g/ml (80%), 1,093 g/ml (75%), 1,087 g/ml (70%), and 1,081 g/ml (65%).

    Techniques: Staining, Incubation