cell toxicity a 549 human lung epithelial cells a 549  (DSMZ)

Journal: International Journal of Molecular Sciences

Article Title: Dual Role of microRNA-146a in Experimental Inflammation in Human Pulmonary Epithelial and Immune Cells and Expression in Inflammatory Lung Diseases

doi: 10.3390/ijms25147686

Figure Lengend Snippet: The in vitro stimulation of lung epithelial cells, macrophages, and neutrophil-precursor cells. Depicted are flow cytometric analyses showing CD69+ ( a – c )- and TLR4+ ( d – f )-stimulated A549, THP1 and HL60 cells. The activation of the NF-kB pathway was detected via a Western blot in all three cell lines, showing the comparison of the NF-kB p50 band in unstimulated and stimulated cells at 50 kDa. YY1 is used as an endogenous control for nuclear protein expression ( g – i ). Further, IL-8 protein expression levels were measured by a cytometric bead array (CBA) after cytokine mix (CM4, CM6, and CM8) stimulation in A549 ( j ) and in HL60 cells ( l ), as well as after M1 differentiation in THP1 cells ( k ). A549: n = 4; THP1: n = 3–8; HL60: n = 3–7. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human lung alveolar epithelial cell line A549 was obtained from DSMZ and was maintained in a DMEM medium (ThermoFisher, Dreieich, Germany) containing 10% fetal calf serum (FCS) (Sigma-Aldrich, Taufkirchen, Germany) and 1% penicillin–streptomycin (Life Technologies, Darmstadt, Germany) at a density of 2 × 10 6 cells per T75 cell culture flask.

Techniques: In Vitro, Activation Assay, Western Blot, Comparison, Control, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Dual Role of microRNA-146a in Experimental Inflammation in Human Pulmonary Epithelial and Immune Cells and Expression in Inflammatory Lung Diseases

doi: 10.3390/ijms25147686

Figure Lengend Snippet: Analysis of miRNA expression in stimulated cell lines. Volcano plots illustrate differentially expressed miRNAs measured by next generation sequencing (NGS) in CM8-stimulated A549 cells ( a ), M1 THP1 cells ( b ), and CM8-stimulated HL60 cells ( c ) compared to unstimulated or M0 THP1 cells, respectively ( n = 4). Green dots represent significantly upregulated miRNAs (log2 fold change (FC) > 1; padj < 0.001) and red dots represent significant downregulation of miRNA expression (log2 FC < −1; padj < 0.001). Vertical dotted lines in the volcano plots demonstrate the thresholds of fold changes for further analyses. Venn blot analysis (Venny 2.1) shows comparison of significantly upregulated miRNAs with padj < 0.001 and log2 FC > 1 in stimulated A549, THP1, and HL60 cells ( d ).

Article Snippet: Human lung alveolar epithelial cell line A549 was obtained from DSMZ and was maintained in a DMEM medium (ThermoFisher, Dreieich, Germany) containing 10% fetal calf serum (FCS) (Sigma-Aldrich, Taufkirchen, Germany) and 1% penicillin–streptomycin (Life Technologies, Darmstadt, Germany) at a density of 2 × 10 6 cells per T75 cell culture flask.

Techniques: Expressing, Next-Generation Sequencing, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Dual Role of microRNA-146a in Experimental Inflammation in Human Pulmonary Epithelial and Immune Cells and Expression in Inflammatory Lung Diseases

doi: 10.3390/ijms25147686

Figure Lengend Snippet: Effects of miRNA transfection on IL-8 mRNA and protein expression. Stimulated A549, THP1, and HL60 cells were transfected with mimics and inhibitors of miR-146a-5p ( a – f ) and miR-146a-3p ( g – l ). IL-8 mRNA expression was measured by qPCR in CM8-stimulated A549 cells ( a , g ), in M1 THP1 cells ( b , h ), and in CM8-stimulated HL60 cells ( c , i ). Protein expression was measured by CBA in supernatants of cell culture samples ( d – f , j – l ). FC = fold change; NC = negative control. A549: n = 6–12, THP1: n = 3–5, HL60: n = 3–6, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human lung alveolar epithelial cell line A549 was obtained from DSMZ and was maintained in a DMEM medium (ThermoFisher, Dreieich, Germany) containing 10% fetal calf serum (FCS) (Sigma-Aldrich, Taufkirchen, Germany) and 1% penicillin–streptomycin (Life Technologies, Darmstadt, Germany) at a density of 2 × 10 6 cells per T75 cell culture flask.

Techniques: Transfection, Expressing, Cell Culture, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: Dual Role of microRNA-146a in Experimental Inflammation in Human Pulmonary Epithelial and Immune Cells and Expression in Inflammatory Lung Diseases

doi: 10.3390/ijms25147686

Figure Lengend Snippet: The impact of the simultaneous modification of miR-146a-5p and miR-146a-3p on cytokine expression. The mimic of miR-146a-5p and the inhibitor of miR-146a-3p were transfected separately and in combination in stimulated A549 ( a , b ) and THP1 cells ( c ), measuring IL-8 ( a , c ) and IL-6 protein expression ( b ) in the supernatant of cell culture samples. The reverse combination consisting of transfection with the miR-146a-5p inhibitor and miR-146a-3p mimics was applied in ( d – f ). A549: n = 5–6, THP1: n = 3; * p < 0.05, ** p < 0.01.

Article Snippet: Human lung alveolar epithelial cell line A549 was obtained from DSMZ and was maintained in a DMEM medium (ThermoFisher, Dreieich, Germany) containing 10% fetal calf serum (FCS) (Sigma-Aldrich, Taufkirchen, Germany) and 1% penicillin–streptomycin (Life Technologies, Darmstadt, Germany) at a density of 2 × 10 6 cells per T75 cell culture flask.

Techniques: Modification, Expressing, Transfection, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Dual Role of microRNA-146a in Experimental Inflammation in Human Pulmonary Epithelial and Immune Cells and Expression in Inflammatory Lung Diseases

doi: 10.3390/ijms25147686

Figure Lengend Snippet: Target analysis of miR-146a-5p and miR-146a-3p. In silico target analysis was performed using www.mirnet.ca showing 203 targets of miR-146a-5p and 116 targets of miR-146a-3p including three overlapping targets. KEGG pathway analysis demonstrates 15 targets implicated in Toll-like receptor signaling (yellow), eight genes involved in RIG-I-like receptor signaling (green), and 12 targets belonging to chemokine signaling pathway (orange) ( a ). Targets that are allocated to multiple pathways are shown as increased dots. Pathways found in the KEGG pathway analysis and their respective padj values are displayed in ( b ) with relevant pathways colored in red. Effects of target modulation by miR-146a-5p mimic and anti-miR-146a-5p transfection were measured individually and in combination in stimulated A549 cells. qPCR measurements show expression of IRAK1 ( c ), TRAF6 ( d ), DDX3X ( e ), RNF125 ( f ), and CXCR4 ( g ) in transfected cells. n = 3. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Human lung alveolar epithelial cell line A549 was obtained from DSMZ and was maintained in a DMEM medium (ThermoFisher, Dreieich, Germany) containing 10% fetal calf serum (FCS) (Sigma-Aldrich, Taufkirchen, Germany) and 1% penicillin–streptomycin (Life Technologies, Darmstadt, Germany) at a density of 2 × 10 6 cells per T75 cell culture flask.

Techniques: In Silico, Transfection, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Dual Role of microRNA-146a in Experimental Inflammation in Human Pulmonary Epithelial and Immune Cells and Expression in Inflammatory Lung Diseases

doi: 10.3390/ijms25147686

Figure Lengend Snippet: Mutual interference of miR-146a-5p and miR-146a-3p expression in vitro. miR-146a-3p expression was measured by qPCR after transfection with miR-146a-5p mimic in stimulated A549 cells ( a ), in M1 THP1 cells ( b ), and in stimulated HL60 cells ( c ). miR-146a-5p expression after transfection with miR-146a-3p mimic in the respective cell lines is depicted in ( d – f ). A549: n = 4–6, THP1: n = 6–7, HL60: n = 3–5. * p < 0.05, **** p < 0.0001.

Article Snippet: Human lung alveolar epithelial cell line A549 was obtained from DSMZ and was maintained in a DMEM medium (ThermoFisher, Dreieich, Germany) containing 10% fetal calf serum (FCS) (Sigma-Aldrich, Taufkirchen, Germany) and 1% penicillin–streptomycin (Life Technologies, Darmstadt, Germany) at a density of 2 × 10 6 cells per T75 cell culture flask.

Techniques: Expressing, In Vitro, Transfection

Journal: Frontiers in Microbiology

Article Title: Silver nanoparticles with plasma-polymerized hexamethyldisiloxane coating on 3D printed substrates are non-cytotoxic and effective against respiratory pathogens

doi: 10.3389/fmicb.2023.1217617

Figure Lengend Snippet: Cytotoxicity of metallic coatings on 3D printed polylactic acid (PLA) substrates on A549 cells seeded directly on the coated carrier (A) or around the coated carrier (B) . Control–uncoated PLA carrier, AgNPs–layer of silver nanoparticles, AgNPs-ppHMDSO–layer of silver nanoparticles overcoated by plasma-polymerized hexamethyldisiloxane (ppHMDSO), Ag–silver coating, Cu–copper coating. Cells were grown for 48 h and their viability was determined using an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) assay. Results are depicted in % of the viability of cells growing on the same area inside a culture plate, three biological replicates ± standard error of the mean.

Article Snippet: Human lung epithelial cells A549 (DSMZ ACC107) were cultured in DMEM (Dulbecco’s Modified Eagle Medium) with 10% FBS (fetal bovine serum) and Pen/Strep (Penicillin Streptomycin).

Techniques: