collagenase d  (Millipore)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Collagenase D
    Description:
    Collagenase D is prepared from Clostridium histolyticum cultures by filtration ammonium sulfate precipitation dialysis and lyophilization
    Catalog Number:
    COLLD-RO
    Price:
    None
    Applications:
    Collagenase from C. histolyticum is widely used for the disaggregation of many types of tissues (e.g., lung, heart, muscle, bone, adipose tissue, liver, kidney, cartilage, mammary gland, placentae, blood vessels, brain, tumors) and for the preparation of single cell suspensions for the establishment of primary cell culture systems. Collagenase D is recommended when functionality and integrity of cell-surface proteins are important.Clostridium collagenase from Roche has been used to prepare cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.
    Buy from Supplier


    Structured Review

    Millipore collagenase d
    Collagenase D
    Collagenase D is prepared from Clostridium histolyticum cultures by filtration ammonium sulfate precipitation dialysis and lyophilization
    https://www.bioz.com/result/collagenase d/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase d - by Bioz Stars, 2021-04
    99/100 stars

    Images

    Related Articles

    Isolation:

    Article Title: Cooperative function of Pdx1 and Oc1 in multipotent pancreatic progenitors impacts postnatal islet maturation and adaptability
    Article Snippet: .. Isolation of islets from P14 pancreas required collagenase digestion of whole dissected pancreas at 37°C and hand-picking of islets away from exocrine tissue following a Histopaque-1077 (Sigma-Aldrich) gradient. ..

    Injection:

    Article Title: Combination of Alphavirus replicon particle-based vaccination with immunomodulatory antibodies: therapeutic activity in the B16 melanoma mouse model and immune correlates
    Article Snippet: .. Sixteen days after injection, matrigel plugs were resected, incubated for 1 hour at 37°C with 1 mg/ml Collagenase D (Sigma) and dissociated to obtain a single-cell suspension. .. Cells were then stained with the following anti-mouse mAbs: anti-CD45.2-APCCY7, anti-CD3-FITC, anti-NK1.1-APC, anti-CD8-PE–Texas Red, anti-CD4-PE, anti-Foxp3-APC, anti-PD-1 APC, anti-OX40-PE, anti-GITR-PECY7, anti-CD27-PE, anti-ICOS-PE (BD Biosciences) and LIVE/DEAD Fixable Aqua Dead Cell Stain kit (ViD, eBiosciences) or DAPI before acquisition.

    Incubation:

    Article Title: Combination of Alphavirus replicon particle-based vaccination with immunomodulatory antibodies: therapeutic activity in the B16 melanoma mouse model and immune correlates
    Article Snippet: .. Sixteen days after injection, matrigel plugs were resected, incubated for 1 hour at 37°C with 1 mg/ml Collagenase D (Sigma) and dissociated to obtain a single-cell suspension. .. Cells were then stained with the following anti-mouse mAbs: anti-CD45.2-APCCY7, anti-CD3-FITC, anti-NK1.1-APC, anti-CD8-PE–Texas Red, anti-CD4-PE, anti-Foxp3-APC, anti-PD-1 APC, anti-OX40-PE, anti-GITR-PECY7, anti-CD27-PE, anti-ICOS-PE (BD Biosciences) and LIVE/DEAD Fixable Aqua Dead Cell Stain kit (ViD, eBiosciences) or DAPI before acquisition.

    Article Title: Polarization of macrophages in the tumor microenvironment is influenced by EGFR signaling within colon cancer cells
    Article Snippet: Flow cytometry Isolation of primary mouse macrophages from colon was performed as described previously [ ]. .. Briefly, colon tissues were cut into small pieces (1-2 mm) and incubated in 10 mL PRMI 1640 medium with 10 mM HEPES and 5% fetal bovine serum containing 10 mg (1 mg/mL) collagenase D (Sigma-Aldrich), 10 mg (1 mg/mL) dispase II (Roche, Germany), and 100 μL 10 mg/ml DNase I (100 μg/mL) (Sigma-Aldrich) for 30-45 min in a shaking incubator at 37°C. .. For Ana-1 and BMDM cells, single-cell suspensions from mouse colon tissues were incubated with APC-anti-mouse CD11b and FITC-anti-mouse F4/80 (Affymetrix eBioscience, USA), or APC-anti-mouse F4/80 and FITC-anti-mouse CD206 (MMR) (Biolegend, San Diego, CA) antibodies.

    Article Title: Resolution of a chronic viral infection after interleukin-10 receptor blockade
    Article Snippet: .. Splenocytes from mice infected 7 d earlier with LCMV Armstrong or LCMV clone 13 with or without anti–IL-10R antibody treatment were incubated with HBSS medium containing 0.5 mg/ml collagenase D (Sigma-Aldrich) at 37°C, 5% CO2 for 30 min. 0.01 M EDTA was added to disrupt T cell–DC complexes. .. Next, cells were depleted of CD3-expressing cells (Dynal CD3-beads; Dynal), incubated with CD11c microbeads (Miltenyi Biotec), and positively selected using MACS columns.

    Flow Cytometry:

    Article Title: Dysregulated Cytokine Expression by CD4+ T cells from Post-Septic Mice Modulates both Th1 and Th2-Mediated Granulomatous Lung Inflammation
    Article Snippet: .. Flow cytometry Collagenase-digested (Sigma-Aldrich, St. Louis, MO) lung lobes and undigested lymph nodes from animals were processed into single cell suspensions by processing tissues through sterile 40-mm filters, and ammonium chloride lysis buffer was used to eliminate erythrocytes. .. Cells were stained with the following fluorescent antibodies in flow cytometry buffer (phosphate buffered saline, 1% w/v bovine serum albumin, 0.05% w/v sodium azide): Purified αCD16/32 (Fc Block) (2.4G2, BD Biosciences, San Jose, CA), FITC-αCD8a (53–6.7, BD Biosciences), FITC-αCD3ε (145-2C11, BD Biosciences), PeCy7-αCD45 (Ly5, eBioscience, San Diego, CA), PeCy7-αCD3ε (17A2, BioLegend, San Diego, CA), Pacific Blue- (RM4-5, BioLegend) or Pacific Orange (RM4-5, Invitrogen, Carlsbad, CA) -αCD4, and Pacific Blue-αCD45 (30-F11, BioLegend).

    Cytometry:

    Article Title: Dysregulated Cytokine Expression by CD4+ T cells from Post-Septic Mice Modulates both Th1 and Th2-Mediated Granulomatous Lung Inflammation
    Article Snippet: .. Flow cytometry Collagenase-digested (Sigma-Aldrich, St. Louis, MO) lung lobes and undigested lymph nodes from animals were processed into single cell suspensions by processing tissues through sterile 40-mm filters, and ammonium chloride lysis buffer was used to eliminate erythrocytes. .. Cells were stained with the following fluorescent antibodies in flow cytometry buffer (phosphate buffered saline, 1% w/v bovine serum albumin, 0.05% w/v sodium azide): Purified αCD16/32 (Fc Block) (2.4G2, BD Biosciences, San Jose, CA), FITC-αCD8a (53–6.7, BD Biosciences), FITC-αCD3ε (145-2C11, BD Biosciences), PeCy7-αCD45 (Ly5, eBioscience, San Diego, CA), PeCy7-αCD3ε (17A2, BioLegend, San Diego, CA), Pacific Blue- (RM4-5, BioLegend) or Pacific Orange (RM4-5, Invitrogen, Carlsbad, CA) -αCD4, and Pacific Blue-αCD45 (30-F11, BioLegend).

    Lysis:

    Article Title: Dysregulated Cytokine Expression by CD4+ T cells from Post-Septic Mice Modulates both Th1 and Th2-Mediated Granulomatous Lung Inflammation
    Article Snippet: .. Flow cytometry Collagenase-digested (Sigma-Aldrich, St. Louis, MO) lung lobes and undigested lymph nodes from animals were processed into single cell suspensions by processing tissues through sterile 40-mm filters, and ammonium chloride lysis buffer was used to eliminate erythrocytes. .. Cells were stained with the following fluorescent antibodies in flow cytometry buffer (phosphate buffered saline, 1% w/v bovine serum albumin, 0.05% w/v sodium azide): Purified αCD16/32 (Fc Block) (2.4G2, BD Biosciences, San Jose, CA), FITC-αCD8a (53–6.7, BD Biosciences), FITC-αCD3ε (145-2C11, BD Biosciences), PeCy7-αCD45 (Ly5, eBioscience, San Diego, CA), PeCy7-αCD3ε (17A2, BioLegend, San Diego, CA), Pacific Blue- (RM4-5, BioLegend) or Pacific Orange (RM4-5, Invitrogen, Carlsbad, CA) -αCD4, and Pacific Blue-αCD45 (30-F11, BioLegend).

    Mouse Assay:

    Article Title: Resolution of a chronic viral infection after interleukin-10 receptor blockade
    Article Snippet: .. Splenocytes from mice infected 7 d earlier with LCMV Armstrong or LCMV clone 13 with or without anti–IL-10R antibody treatment were incubated with HBSS medium containing 0.5 mg/ml collagenase D (Sigma-Aldrich) at 37°C, 5% CO2 for 30 min. 0.01 M EDTA was added to disrupt T cell–DC complexes. .. Next, cells were depleted of CD3-expressing cells (Dynal CD3-beads; Dynal), incubated with CD11c microbeads (Miltenyi Biotec), and positively selected using MACS columns.

    Infection:

    Article Title: Resolution of a chronic viral infection after interleukin-10 receptor blockade
    Article Snippet: .. Splenocytes from mice infected 7 d earlier with LCMV Armstrong or LCMV clone 13 with or without anti–IL-10R antibody treatment were incubated with HBSS medium containing 0.5 mg/ml collagenase D (Sigma-Aldrich) at 37°C, 5% CO2 for 30 min. 0.01 M EDTA was added to disrupt T cell–DC complexes. .. Next, cells were depleted of CD3-expressing cells (Dynal CD3-beads; Dynal), incubated with CD11c microbeads (Miltenyi Biotec), and positively selected using MACS columns.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    Millipore p14 pancreas required collagenase digestion
    Double heterozygotes have significant alterations in postnatal islet gene expression. A : expression of select endocrine development and functional genes in <t>P14</t> islets represented in a heatmap. Green, increased gene expression; red, reduced gene expression. Blue boxes, controls; green boxes, PH; red boxes, OH; black boxes, DH. B – D : relative expression levels of hormones ( B ), regulatory transcription factors ( C ), and islet functional genes ( D ) ( n = 4). One symbol, P ≤ 0.05; two symbols, P ≤ 0.01; three symbols, P ≤ 0.001. +Control (Con) vs. PH; #DH vs. PH; ^OH vs. DH; *Con vs. DH by 2-way ANOVA with Tukey’s correction for multiple comparisons or Kruskal-Wallis with Dunn’s correction for multiple comparisons.
    P14 Pancreas Required Collagenase Digestion, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p14 pancreas required collagenase digestion/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p14 pancreas required collagenase digestion - by Bioz Stars, 2021-04
    97/100 stars
      Buy from Supplier

    97
    Millipore control collagenase inhibitor z pro d leu d ala nhoh
    Neutrophil elastase potentiates the microbicidal response of T. cruzi in vitro . (A) Parasites released in infected C57BL/6 macrophages cultures alone or in presence of solvent (DMSO), control inhibitor (collagenase inhibitor <t>Z-Pro-D-Leu-AAPV-D-Ala-NHOH;</t> used at 10 µg/ml) or NE inhibitor (MeOSuc-AAPV-cmk; used at 10 µg/ml). (B) Neutrophil elastase (100 ng/ml) was added to infected cultured macrophages in the presence or absence of selective iNOS inhibitor (L-NIL). The number of trypomastigotes was counted in a Neubauer chamber after 7 days of culture. Production of TNF-α and NO released into the supernatant by T. cruzi infected macrophages after 24 h incubation in medium alone or in the presence of neutrophil elastase, at 100 ng/ml. Measurements were performed in triplicates of three different experiments. Statistical significance was determined by ANOVA (A) or t test (B).
    Control Collagenase Inhibitor Z Pro D Leu D Ala Nhoh, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/control collagenase inhibitor z pro d leu d ala nhoh/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    control collagenase inhibitor z pro d leu d ala nhoh - by Bioz Stars, 2021-04
    97/100 stars
      Buy from Supplier

    Image Search Results


    Double heterozygotes have significant alterations in postnatal islet gene expression. A : expression of select endocrine development and functional genes in P14 islets represented in a heatmap. Green, increased gene expression; red, reduced gene expression. Blue boxes, controls; green boxes, PH; red boxes, OH; black boxes, DH. B – D : relative expression levels of hormones ( B ), regulatory transcription factors ( C ), and islet functional genes ( D ) ( n = 4). One symbol, P ≤ 0.05; two symbols, P ≤ 0.01; three symbols, P ≤ 0.001. +Control (Con) vs. PH; #DH vs. PH; ^OH vs. DH; *Con vs. DH by 2-way ANOVA with Tukey’s correction for multiple comparisons or Kruskal-Wallis with Dunn’s correction for multiple comparisons.

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: Cooperative function of Pdx1 and Oc1 in multipotent pancreatic progenitors impacts postnatal islet maturation and adaptability

    doi: 10.1152/ajpendo.00260.2017

    Figure Lengend Snippet: Double heterozygotes have significant alterations in postnatal islet gene expression. A : expression of select endocrine development and functional genes in P14 islets represented in a heatmap. Green, increased gene expression; red, reduced gene expression. Blue boxes, controls; green boxes, PH; red boxes, OH; black boxes, DH. B – D : relative expression levels of hormones ( B ), regulatory transcription factors ( C ), and islet functional genes ( D ) ( n = 4). One symbol, P ≤ 0.05; two symbols, P ≤ 0.01; three symbols, P ≤ 0.001. +Control (Con) vs. PH; #DH vs. PH; ^OH vs. DH; *Con vs. DH by 2-way ANOVA with Tukey’s correction for multiple comparisons or Kruskal-Wallis with Dunn’s correction for multiple comparisons.

    Article Snippet: Isolation of islets from P14 pancreas required collagenase digestion of whole dissected pancreas at 37°C and hand-picking of islets away from exocrine tissue following a Histopaque-1077 (Sigma-Aldrich) gradient.

    Techniques: Expressing, Functional Assay

    Islets of double heterozygotes show altered postnatal maturation. A : P14 DH islets lack a distinct β-cell core and α-cell mantle (green, insulin; red, glucagon; blue, DAPI). B – D : quantification of the percentage of islets with a mixed phenotype ( B ) and the percentage of nonmantle α-cells in each genotype at P14 ( C ), without a change in β- to α-cell ratio ( D ) ( n = 5–7). E and F : urocortin3 mRNA ( E ) and protein ( F ) are reduced in PH and DH islets ( n = 3 for mRNA, images representative of 6 animals). Scale bar: 100 μm. * P ≤ 0.05.

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: Cooperative function of Pdx1 and Oc1 in multipotent pancreatic progenitors impacts postnatal islet maturation and adaptability

    doi: 10.1152/ajpendo.00260.2017

    Figure Lengend Snippet: Islets of double heterozygotes show altered postnatal maturation. A : P14 DH islets lack a distinct β-cell core and α-cell mantle (green, insulin; red, glucagon; blue, DAPI). B – D : quantification of the percentage of islets with a mixed phenotype ( B ) and the percentage of nonmantle α-cells in each genotype at P14 ( C ), without a change in β- to α-cell ratio ( D ) ( n = 5–7). E and F : urocortin3 mRNA ( E ) and protein ( F ) are reduced in PH and DH islets ( n = 3 for mRNA, images representative of 6 animals). Scale bar: 100 μm. * P ≤ 0.05.

    Article Snippet: Isolation of islets from P14 pancreas required collagenase digestion of whole dissected pancreas at 37°C and hand-picking of islets away from exocrine tissue following a Histopaque-1077 (Sigma-Aldrich) gradient.

    Techniques:

    Double heterozygosity negatively impacts expression of regulatory and functional factors in postnatal islets. A : Nkx6.1 (red) is detected in fewer β-cells (insulin, green) and has reduced intensity in P14 DH. Arrowheads in DH indicate Nkx6.1 + cells. B : quantification of A ( n = 4–6). C : Glut2 expression (red) is weaker and more diffuse (E-cadherin, green) in β-cells (insulin, yellow) of DH. DAPI is in blue ( n = 4–6). Scale bar: 100 μm. **** P ≤ 0.0001 by 1-Way ANOVA with Tukey’s correction for multiple comparisons.

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: Cooperative function of Pdx1 and Oc1 in multipotent pancreatic progenitors impacts postnatal islet maturation and adaptability

    doi: 10.1152/ajpendo.00260.2017

    Figure Lengend Snippet: Double heterozygosity negatively impacts expression of regulatory and functional factors in postnatal islets. A : Nkx6.1 (red) is detected in fewer β-cells (insulin, green) and has reduced intensity in P14 DH. Arrowheads in DH indicate Nkx6.1 + cells. B : quantification of A ( n = 4–6). C : Glut2 expression (red) is weaker and more diffuse (E-cadherin, green) in β-cells (insulin, yellow) of DH. DAPI is in blue ( n = 4–6). Scale bar: 100 μm. **** P ≤ 0.0001 by 1-Way ANOVA with Tukey’s correction for multiple comparisons.

    Article Snippet: Isolation of islets from P14 pancreas required collagenase digestion of whole dissected pancreas at 37°C and hand-picking of islets away from exocrine tissue following a Histopaque-1077 (Sigma-Aldrich) gradient.

    Techniques: Expressing, Functional Assay

    Anti–IL-10R treatment restores cell numbers in LCMV clone 13–infected mice and resets the CD8α −  versus CD8α +  DC ratio to levels observed in LCMV Armstrong–infected mice.  (A) The effect of 250 μg anti–IL-10R antibody (i.p.) or IgG 1  isotype control antibody treatment on different splenic cell subsets was monitored quantitatively. Spleens of mice infected 3 or 21 wk earlier with LCMV clone 13 were harvested, and cell suspensions were stained with fluorescent antibodies to CD4, CD8, B220, CD11c, and CD11b. Age-matched naive mice were included as controls. Numbers of cells were quantified by relating the percentage to total numbers of spleen cells. Mean values for three to five individual mice are shown. Results are representative of three similar experiments. (B and C) The total numbers of CD11c + CD8α −  (B, top) or CD11c + CD8α +  splenic DC subsets (B, bottom) and the CD8α −  to CD8α +  DC ratios (C) were determined 0, 1, 2, and 6 wk after LCMV Armstrong or LCMV clone 13 infection. In addition, absolute splenic DC subsets (B) and the CD8α −  to CD8α +  DC ratio (C) were calculated from mice therapeutically treated with anti–IL10R on days 7 and 14 after LCMV clone 13 infection; the latter was examined at 6 wk after infection only. Cell suspensions were prepared by digestion with collagenase D. Cells were then stained with fluorescent antibodies to CD11c, CD3, and CD8α. The percentage of CD8α −  and CD8α +  within the CD11c + CD3 −  subset was determined by flow cytometry, and total numbers per spleen were quantified by relating the percentage of these cells to total numbers of CD11c + CD3 −  spleen cells. Histogram bars represent mean values ± SD for three mice per group. The experiment is representative of three similar experiments. Statistical analysis was performed using the Student's  t  test. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: Resolution of a chronic viral infection after interleukin-10 receptor blockade

    doi: 10.1084/jem.20061462

    Figure Lengend Snippet: Anti–IL-10R treatment restores cell numbers in LCMV clone 13–infected mice and resets the CD8α − versus CD8α + DC ratio to levels observed in LCMV Armstrong–infected mice. (A) The effect of 250 μg anti–IL-10R antibody (i.p.) or IgG 1 isotype control antibody treatment on different splenic cell subsets was monitored quantitatively. Spleens of mice infected 3 or 21 wk earlier with LCMV clone 13 were harvested, and cell suspensions were stained with fluorescent antibodies to CD4, CD8, B220, CD11c, and CD11b. Age-matched naive mice were included as controls. Numbers of cells were quantified by relating the percentage to total numbers of spleen cells. Mean values for three to five individual mice are shown. Results are representative of three similar experiments. (B and C) The total numbers of CD11c + CD8α − (B, top) or CD11c + CD8α + splenic DC subsets (B, bottom) and the CD8α − to CD8α + DC ratios (C) were determined 0, 1, 2, and 6 wk after LCMV Armstrong or LCMV clone 13 infection. In addition, absolute splenic DC subsets (B) and the CD8α − to CD8α + DC ratio (C) were calculated from mice therapeutically treated with anti–IL10R on days 7 and 14 after LCMV clone 13 infection; the latter was examined at 6 wk after infection only. Cell suspensions were prepared by digestion with collagenase D. Cells were then stained with fluorescent antibodies to CD11c, CD3, and CD8α. The percentage of CD8α − and CD8α + within the CD11c + CD3 − subset was determined by flow cytometry, and total numbers per spleen were quantified by relating the percentage of these cells to total numbers of CD11c + CD3 − spleen cells. Histogram bars represent mean values ± SD for three mice per group. The experiment is representative of three similar experiments. Statistical analysis was performed using the Student's t test. *, P

    Article Snippet: Splenocytes from mice infected 7 d earlier with LCMV Armstrong or LCMV clone 13 with or without anti–IL-10R antibody treatment were incubated with HBSS medium containing 0.5 mg/ml collagenase D (Sigma-Aldrich) at 37°C, 5% CO2 for 30 min. 0.01 M EDTA was added to disrupt T cell–DC complexes.

    Techniques: Infection, Mouse Assay, Staining, Flow Cytometry, Cytometry

    Spreading of IFNγ signaling in tumor xenografts treated in vivo with collagenase D. a CXCL9 fold induction along the tumor sections in mice treated with IFNγ alone (50 ng per tumor) or together with collagenase D (2.5 μg per tumor). Mean ± SEM of six independent tumors for each treatment. For CXCL9 induction in individual tumors see Supplementary Fig. 6b . *** P = 0.0006; Paired t -test, t = 4.035 and degree of freedom = 20. The red dotted line marks the threshold of 50 CXCL9 mRNA copies per section of the tumor. Each section represents 0.66 mm thick, contains 30 tumor slices, and about 1–4 millions cells. The drawing below shows the diffusion of CXCL9 gradient along the tumor taking into account the threshold. b Quantification of CXCL9 staining area in immunohistochemistry images of several sections of tumors treated either with IFNγ alone or combined with collagenase D. Representative images are shown in Supplementary Fig. 8

    Journal: Nature Communications

    Article Title: Galectin-3 captures interferon-gamma in the tumor matrix reducing chemokine gradient production and T-cell tumor infiltration

    doi: 10.1038/s41467-017-00925-6

    Figure Lengend Snippet: Spreading of IFNγ signaling in tumor xenografts treated in vivo with collagenase D. a CXCL9 fold induction along the tumor sections in mice treated with IFNγ alone (50 ng per tumor) or together with collagenase D (2.5 μg per tumor). Mean ± SEM of six independent tumors for each treatment. For CXCL9 induction in individual tumors see Supplementary Fig. 6b . *** P = 0.0006; Paired t -test, t = 4.035 and degree of freedom = 20. The red dotted line marks the threshold of 50 CXCL9 mRNA copies per section of the tumor. Each section represents 0.66 mm thick, contains 30 tumor slices, and about 1–4 millions cells. The drawing below shows the diffusion of CXCL9 gradient along the tumor taking into account the threshold. b Quantification of CXCL9 staining area in immunohistochemistry images of several sections of tumors treated either with IFNγ alone or combined with collagenase D. Representative images are shown in Supplementary Fig. 8

    Article Snippet: Collagenase D from Clostridium Histolyticum was obtained from Sigma-Aldrich (#11088858001).

    Techniques: In Vivo, Mouse Assay, Diffusion-based Assay, Staining, Immunohistochemistry

    Neutrophil elastase potentiates the microbicidal response of T. cruzi in vitro . (A) Parasites released in infected C57BL/6 macrophages cultures alone or in presence of solvent (DMSO), control inhibitor (collagenase inhibitor Z-Pro-D-Leu-AAPV-D-Ala-NHOH; used at 10 µg/ml) or NE inhibitor (MeOSuc-AAPV-cmk; used at 10 µg/ml). (B) Neutrophil elastase (100 ng/ml) was added to infected cultured macrophages in the presence or absence of selective iNOS inhibitor (L-NIL). The number of trypomastigotes was counted in a Neubauer chamber after 7 days of culture. Production of TNF-α and NO released into the supernatant by T. cruzi infected macrophages after 24 h incubation in medium alone or in the presence of neutrophil elastase, at 100 ng/ml. Measurements were performed in triplicates of three different experiments. Statistical significance was determined by ANOVA (A) or t test (B).

    Journal: PLoS ONE

    Article Title: Neutrophils Increase or Reduce Parasite Burden in Trypanosoma cruzi-Infected Macrophages, Depending on Host Strain: Role of Neutrophil Elastase

    doi: 10.1371/journal.pone.0090582

    Figure Lengend Snippet: Neutrophil elastase potentiates the microbicidal response of T. cruzi in vitro . (A) Parasites released in infected C57BL/6 macrophages cultures alone or in presence of solvent (DMSO), control inhibitor (collagenase inhibitor Z-Pro-D-Leu-AAPV-D-Ala-NHOH; used at 10 µg/ml) or NE inhibitor (MeOSuc-AAPV-cmk; used at 10 µg/ml). (B) Neutrophil elastase (100 ng/ml) was added to infected cultured macrophages in the presence or absence of selective iNOS inhibitor (L-NIL). The number of trypomastigotes was counted in a Neubauer chamber after 7 days of culture. Production of TNF-α and NO released into the supernatant by T. cruzi infected macrophages after 24 h incubation in medium alone or in the presence of neutrophil elastase, at 100 ng/ml. Measurements were performed in triplicates of three different experiments. Statistical significance was determined by ANOVA (A) or t test (B).

    Article Snippet: Monolayers were also treated with the specific NE inhibitor methoxysuccinyl-Ala-Ala-Pro-Val-chloromethylketone (MeoSuc-AAPV-CMK; Calbiochen-Novabiochem, La Jolla, CA), control collagenase inhibitor Z-Pro-D-Leu-D-Ala-NHOH (Calbiochem-Novabiochem), both at 10 µg/ml, or equivalent dosage of solvent (DMSO) alone.

    Techniques: In Vitro, Infection, Cell Culture, Incubation