Journal: Cell Proliferation

Article Title: Isolation and characterization of multipotent stem cells from human cruciate ligaments

doi: 10.1111/j.1365-2184.2009.00611.x

Figure Lengend Snippet: Immunophenotype of ligament‐derived stem cells (LSC) isolated from anterior (ACL) and posterior cruciate ligament (PCL). LSCs from ACL and PCL were cultured for 3–5 passages, harvested, and labelled with antibodies against human antigens CD13, CD14, CD29, CD34, CD44, CD45, CD73, CD90, CD105, CD133, CD166, HLAABC, and HLA‐DR, as indicated and analysed by FACS. (a) LSCs from ACL. (b) LSCs from PCL.

Article Snippet: Antibodies against human CD13, CD14, CD29, CD34, CD44, CD45, CD73, CD90, CD105, CD166, HLA‐ABC and HLA‐DR were purchased from Becton Dickinson; antibodies against human CD133 were purchased from Miltenyi Biotec (Bergisch Glabach, Germany).

Techniques: Derivative Assay, Isolation, Cell Culture

Journal: Cell Proliferation

Article Title: Isolation and characterization of multipotent stem cells from human cruciate ligaments

doi: 10.1111/j.1365-2184.2009.00611.x

Figure Lengend Snippet: The average values of surface immunophenotypes of ligament‐derived stem cells (LSC) from anterior (ACL) and posterior cruciate ligament (PCL) of three different donors

Article Snippet: Antibodies against human CD13, CD14, CD29, CD34, CD44, CD45, CD73, CD90, CD105, CD166, HLA‐ABC and HLA‐DR were purchased from Becton Dickinson; antibodies against human CD133 were purchased from Miltenyi Biotec (Bergisch Glabach, Germany).

Techniques:

Journal: PLoS ONE

Article Title: Platelet Surface-Associated Activation and Secretion-Mediated Inhibition of Coagulation Factor XII

doi: 10.1371/journal.pone.0116665

Figure Lengend Snippet: Stimulation and platelet-dependent fXII activation.

Article Snippet: The following materials were used: thrombin (Haematologic Technologies; Essex Junction, VT, USA); fXIIa and fXII (Enzyme Research Laboratories; South Bend, IN, USA); AlignFlow flow cytometry alignment beads (2.5 μm for 488-nm excitation), fluorescein-5-isothiocyanate (FITC), and tetramethylrhodamine (TMRM) (Molecular Probes; Eugene, OR, USA); unlabelled and FITC-annexin V (BD Biosciences; San Jose, CA, USA); AlexaFluor-647-annexin V (Biolegend; San Diego, CA, USA); prostaglandin E1 (PGE1) (MP Biochemicals; Irvine, CA, USA); PPACK (EMD Chemicals; Gibbstown, NJ, USA); the chromomeric substrates S2238 and S2302 (Chromogenix; Milano, Italy); HEPES, bovine serum albumin, Sepharose CL-2B, Protein G sepharose, apyrase grade VII, mepacrine (quinacrine), PBS, EDTA and DMSO (Sigma-Aldrich; St Louis, MO, USA); calpeptin and MDL 28170 (Tocris Bioscience; Ellisville, MO, USA); G1/C1-inhibitor and polyclonal goat anti-human serpin G1/C1-inhibitor antibody (R&D Systems; Minneapolis, MN, USA); goat polyclonal anti-human factor XII antibody (LifeSpan BioSciences, Inc., Seattle, WA, USA); peroxidase-AffiniPure donkey anti-goat IgG (Jackson ImmunoResearch Laboratories; West Grove, PA, USA); egg phosphatidylcholine and egg phosphatidylserine (Avanti Polar Lipids; Alabaster, AL, USA); DAPI (4′,6-Diamidino-2-phenylindole dihydrochloride) (AppliChem; Darmstadt, Germany); and a kit to estimate fXII activation (Renam; Moscow, Russia).

Techniques: Activation Assay, Significance Assay

Journal: PLoS ONE

Article Title: Platelet Surface-Associated Activation and Secretion-Mediated Inhibition of Coagulation Factor XII

doi: 10.1371/journal.pone.0116665

Figure Lengend Snippet: ( A ) We used a scheme that depicts the platelet aggregate to test the C1-INH washout hypothesis. The platelet concentration inside the thrombi was considered 1 per 15 fl. The initial C1-inhibitor distribution in the aggregate for the simulations is shown in grey, and at values ranging from 0.1 to 10 μm/s, the flow penetrated the aggregate. At t = 0, C1-INH at 100 μM (estimated from ref. ) and fXII at 450 nM appeared simultaneously. Factor XII could be activated on the platelet surface, and C1-INH could diffuse through the aggregate (D = 10 μm 2 /s, based on the molecular weight) and move due to the flow. The fXIIa diffusion was assumed negligible because fXII activation is surface-associated ( and this study), and typically, fXIIa is tightly bound to the activation surface . The C1-INH action was described using a mass action equation with the reaction constant 0.00366 μM -1 s -1 . ( B ) Time-course of the distance-averaged [C1-INH] for various flow velocities. ( C ) Time-course of the surface-averaged [fXIIa] for various flow velocities. The C1-INH and fXIIa spatial distributions were governed by a set of differential equations, which were solved using the finite volume solver available within the Virtual Cell environment.

Article Snippet: The following materials were used: thrombin (Haematologic Technologies; Essex Junction, VT, USA); fXIIa and fXII (Enzyme Research Laboratories; South Bend, IN, USA); AlignFlow flow cytometry alignment beads (2.5 μm for 488-nm excitation), fluorescein-5-isothiocyanate (FITC), and tetramethylrhodamine (TMRM) (Molecular Probes; Eugene, OR, USA); unlabelled and FITC-annexin V (BD Biosciences; San Jose, CA, USA); AlexaFluor-647-annexin V (Biolegend; San Diego, CA, USA); prostaglandin E1 (PGE1) (MP Biochemicals; Irvine, CA, USA); PPACK (EMD Chemicals; Gibbstown, NJ, USA); the chromomeric substrates S2238 and S2302 (Chromogenix; Milano, Italy); HEPES, bovine serum albumin, Sepharose CL-2B, Protein G sepharose, apyrase grade VII, mepacrine (quinacrine), PBS, EDTA and DMSO (Sigma-Aldrich; St Louis, MO, USA); calpeptin and MDL 28170 (Tocris Bioscience; Ellisville, MO, USA); G1/C1-inhibitor and polyclonal goat anti-human serpin G1/C1-inhibitor antibody (R&D Systems; Minneapolis, MN, USA); goat polyclonal anti-human factor XII antibody (LifeSpan BioSciences, Inc., Seattle, WA, USA); peroxidase-AffiniPure donkey anti-goat IgG (Jackson ImmunoResearch Laboratories; West Grove, PA, USA); egg phosphatidylcholine and egg phosphatidylserine (Avanti Polar Lipids; Alabaster, AL, USA); DAPI (4′,6-Diamidino-2-phenylindole dihydrochloride) (AppliChem; Darmstadt, Germany); and a kit to estimate fXII activation (Renam; Moscow, Russia).

Techniques: Concentration Assay, Molecular Weight, Diffusion-based Assay, Activation Assay

Journal:

Article Title: Involvement of Aminopeptidase N (CD13) in Infection of Human Neural Cells by Human Coronavirus 229E

doi:

Figure Lengend Snippet: Flow cytometry analysis of cell surface expression of CD13. Cells were labeled with either an anti-CD13 PE-conjugated antibody or a PE-conjugated isotypic control antibody. (A) THP-1 monocytic cell line (2-log-unit signal displacement); (B) L-132 human lung fibroblast cell line (1-log-unit signal displacement); (C) CHME-5 microglial cell line (no signal displacement); (D) H4 neuronal cell line (1-log-unit signal displacement); (E) SK-N-SH neuronal cell line (2-log-unit signal displacement); (F) MO3.13 oligodendrocytic cell line (0.2-log-unit signal displacement); (G) GL-15 astrocytic cell line (0.5-log-unit signal displacement); (H) U-87 MG astrocytic cell line (0.1-log-unit signal displacement); (I) U-373 MG astrocytic cell line (0.1-log-unit signal displacement). These profiles are representative of at least two separate experiments.

Article Snippet: The absence of interaction between the anti-CD13 MAb (azide-free WM15; Biodesign International, Kennebunk, Maine) and HCV-229E was determined by an enzyme-linked immunosorbent assay (ELISA).

Techniques: Flow Cytometry, Expressing, Labeling

Journal:

Article Title: Involvement of Aminopeptidase N (CD13) in Infection of Human Neural Cells by Human Coronavirus 229E

doi:

Figure Lengend Snippet: Detection of binding of 35S-labeled HCV-229E on cultures of human neural cell lines pretreated with an anti-CD13 MAb or with various controls, all in triplicate. (A) L-132 human lung fibroblast cell line; (B) CHME-5 microglial cell line; (C) H4 neuronal cell line; (D) SK-N-SH neuronal cell line; (E) MO3.13 oligodendrocytic cell line; (F) GL-15 astrocytic cell line; (G) U-373 MG astrocytic cell line; (H) U-87 MG astrocytic cell line. Treatment 1, culture medium; treatment 2, 105 dpm of HCV-229E; treatment 3, 105 dpm of HCV-229E plus HLA-specific antiserum (15 μg); treatment 4, 105 dpm of HCV-229E plus anti-TuMV MAb 6D (15 μg); treatment 5, 105 dpm of HCV-229E plus anti-HCV-229E MAb 5-11H.6 (15 μg); treatment 6, 105 dpm of HCV-229E plus anti-CD13 MAb WM15 (15 μg); treatment 7, 105 dpm of HCV-229E plus unlabeled HCV-229E. These profiles are representative of two separate experiments. ∗∗, statistically significant difference according to Tukey’s multiple comparison test, following an ANOVA test (P < 0.05).

Article Snippet: The absence of interaction between the anti-CD13 MAb (azide-free WM15; Biodesign International, Kennebunk, Maine) and HCV-229E was determined by an enzyme-linked immunosorbent assay (ELISA).

Techniques: Binding Assay, Labeling

Journal:

Article Title: Involvement of Aminopeptidase N (CD13) in Infection of Human Neural Cells by Human Coronavirus 229E

doi:

Figure Lengend Snippet: Correlation between expression levels of CD13 on human neural cell lines and attachment of 35S-labeled HCV-229E. ▪, L-132 human lung fibroblastic cell line; ▵, H4 neuronal cell line; ▴, SK-N-SH neuronal cell line; ⧫, GL-15 astrocytic cell line; ○, U-87 MG astrocytic cell line; ◊, U-373 MG astrocytic cell line; •, MO3.13 oligodendrocytic cell line; □, CHME-5 microglial cell line.

Article Snippet: The absence of interaction between the anti-CD13 MAb (azide-free WM15; Biodesign International, Kennebunk, Maine) and HCV-229E was determined by an enzyme-linked immunosorbent assay (ELISA).

Techniques: Expressing, Labeling

Journal:

Article Title: Involvement of Aminopeptidase N (CD13) in Infection of Human Neural Cells by Human Coronavirus 229E

doi:

Figure Lengend Snippet: Detection of viral antigens by indirect immunofluorescence in cultures of human neural cells pretreated with an anti-CD13 MAb and inoculated with HCV-229E. (A and B) L-132 human lung fibroblastic cell line; (C and D) SK-N-SH neuronal cell line; (E and F) GL-15 astrocytic cell line; (G and H) MO3.13 oligodendrocytic cell line. Cells were pretreated with 15 μg of an anti-CD13 MAb (B, D, F, and H) or with 15 μg of an isotypic control antibody (A, C, E, and G) and were inoculated with HCV-229E at an MOI of 0.1. After 22 h, the cells were fixed in cold acetone and examined by indirect immunofluorescence. The primary antibody used to detect viral antigens was a guinea pig antiserum against HCV-229E. This was followed by the addition of a fluorescein-conjugated goat affinity-purified anti-guinea pig secondary antibody Magnification, ×100. The results represent several fields, and observations at lower magnification yielded the following percentages of infected cells and qualitative levels of fluorescence (scale, 0 to 3+): 30% and 3+ (A and C), 10% and 1.5+ (E), 10 to 15% and 3+ (G), <5% and 1+ (B), <5% and 0.5+ (D and F), and negative (H).

Article Snippet: The absence of interaction between the anti-CD13 MAb (azide-free WM15; Biodesign International, Kennebunk, Maine) and HCV-229E was determined by an enzyme-linked immunosorbent assay (ELISA).

Techniques: Immunofluorescence, Affinity Purification, Infection, Fluorescence