Journal: Frontiers in Immunology

Article Title: The mammosphere-derived epithelial cell secretome modulates neutrophil functions in the bovine model

doi: 10.3389/fimmu.2024.1367432

Figure Lengend Snippet: Primers used for reverse transcriptase (RT)-PCR.

Article Snippet: Treatments consisting of either RPMI + 2% FBS (control), MDEC CM, 20 ng/ml bovine granulocyte-macrophage colony-stimulating factor (GM CSF) or 50 ng/ml bovine insulin-like growth factor 2 (IGF2) (both from Kingfisher Biotech, and diluted in RPMI + 2% FBS), were added, and cells were incubated for 1 h at 37°C.

Techniques: Reverse Transcription

Journal: Frontiers in Immunology

Article Title: The mammosphere-derived epithelial cell secretome modulates neutrophil functions in the bovine model

doi: 10.3389/fimmu.2024.1367432

Figure Lengend Snippet: Proteins in bovine mammosphere-derived epithelial cell (MDEC) conditioned medium (CM) affecting neutrophil functions, as detected by liquid chromatography-mass spectrometry.

Article Snippet: Treatments consisting of either RPMI + 2% FBS (control), MDEC CM, 20 ng/ml bovine granulocyte-macrophage colony-stimulating factor (GM CSF) or 50 ng/ml bovine insulin-like growth factor 2 (IGF2) (both from Kingfisher Biotech, and diluted in RPMI + 2% FBS), were added, and cells were incubated for 1 h at 37°C.

Techniques: Chromatography

Journal: Frontiers in Immunology

Article Title: The mammosphere-derived epithelial cell secretome modulates neutrophil functions in the bovine model

doi: 10.3389/fimmu.2024.1367432

Figure Lengend Snippet: Reducing Chemokine (C-X-C motif) ligand 6 (CXCL6) expression in bovine mammosphere-derived epithelial cells (MDECs) changes the effect of conditioned medium (CM) on neutrophil chemotaxis, while reducing insulin-like growth factor 2 (IGF2) expression in MDECs does not change the effect of conditioned medium (CM) on neutrophil phagocytosis. (A) . Recombinant bovine CXCL6 was added to bovine neutrophils to determine if chemotaxis changed relative to RPMI control medium (dotted line). Cells treated with MDEC CM were included as a control. (B) . CXCL6 siRNA was used to knock down CXCL6 in MDEC CM. Chemotaxis was assessed in neutrophils treated with RPMI control medium, MDEC CM, MDEC CM from cells transfected with CXCL6 siRNA, and MDEC CM from cells transfected with a non-specific (negative) siRNA. (C) . Recombinant bovine IGF2 was added to bovine neutrophils to determine if phagocytosis changed relative to the RPMI control (dotted line). Cells treated with MDEC CM were included as a control. (D) . IGF2 siRNA was used to knock down IGF2 in MDEC CM. Phagocytosis was assessed in neutrophils treated with RPMI control medium, MDEC CM, MDEC CM from cells transfected with IGF2 siRNA and MDEC CM from cells that received a non-specific (negative) siRNA. Each data point on graphs represents the results from one experiment, assessing the effects of CM from one MDEC line on 3 neutrophil preparations. Gray dotted lines on graphs indicate the control conditions expressed as 100%. n = 3 experiments.

Article Snippet: Treatments consisting of either RPMI + 2% FBS (control), MDEC CM, 20 ng/ml bovine granulocyte-macrophage colony-stimulating factor (GM CSF) or 50 ng/ml bovine insulin-like growth factor 2 (IGF2) (both from Kingfisher Biotech, and diluted in RPMI + 2% FBS), were added, and cells were incubated for 1 h at 37°C.

Techniques: Expressing, Derivative Assay, Chemotaxis Assay, Recombinant, Control, Knockdown, Transfection