human telomerase-immortalized coronary artery endothelial (ticae) cells  (European Collection of Authenticated Cell Cultures)

Journal: Frontiers in Pharmacology

Article Title: Connexin43 Hemichannel Targeting With TAT-Gap19 Alleviates Radiation-Induced Endothelial Cell Damage

doi: 10.3389/fphar.2020.00212

Figure Lengend Snippet: Radiation-induced inflammatory responses and the effect of TAT-Gap19. The response of various inflammatory markers in (A–H) TICAE cells and (I–X) TIME cells to 0.1 and 5 Gy irradiation conditions. The values represent the average ± SEM of 5–6 biological replicates; data were analyzed with a non-parametric Mann–Whitney T -test. *Indicates statistical differences compared to the respective 0 Gy controls. •Indicates statistically significant differences between the TAT-Gap19 group compared to the corresponding responses in the control group (not-treated with TAT-Gap19). */• p < 0.05; **/•• p < 0.01. IL-1β, Interleukin 1 beta; IL-6, Interleukin 6; IL-8, Interleukin 8; ICAM-1, Intracellular adhesion molecule 1; VCAM-1, Vascular cell adhesion protein 1; PECAM-1, Platelet endothelial cell adhesion molecule-1; MCP-1, Monocyte chemotactic protein-1; TNF- α, Tumor necrosis factor alpha; CRP, C-reactive protein.

Article Snippet: Two human endothelial cell lines: Telomerase Immortalized human Coronary Artery Endothelial cells (TICAE) from the European Collection of Authenticated Cell Cultures (ECACC), and Telomerase Immortalized human Microvascular Endothelial cells (TIME) from the American Type Cell Culture (ATTC), were grown in MesoEndo Cell Growth Medium (Sigma-Aldrich Co., LCC, Diegem, Belgium) at 37°C in a humidified incubator supplemented with 5% CO2.

Techniques: Irradiation, MANN-WHITNEY, Control

Journal: Frontiers in Pharmacology

Article Title: Connexin43 Hemichannel Targeting With TAT-Gap19 Alleviates Radiation-Induced Endothelial Cell Damage

doi: 10.3389/fphar.2020.00212

Figure Lengend Snippet: Radiation-induced premature endothelial senescence and effect of TAT-Gap19. Senescence-associated β-galactosidase activity was measured in a CPRG assay at 7, 9, and 14 days after radiation exposure (0.1 and 5 Gy) in (A) TICAE cells and (B) TIME cells. IGFBP-7 and GDF-15 were assessed in (C) TICAE cells and (D) TIME cells at 7 days p.i. The values represent the average ± SEM of 16–24 biological replicates in the CPRG assay and of six biological replicates in multiplex-based assays. Statistical significance was analyzed with a non-parametric Mann–Whitney T -test. *Indicates statistically significant differences compared to the respective 0 Gy controls. •Indicates statistically significant differences between the TAT-Gap19 group compared to the respective control condition (not treated with TAT-Gap19). */• p < 0.05; **/•• p < 0.01., ***/••• P < 0.001, ****/•••• P < 0.0001.

Article Snippet: Two human endothelial cell lines: Telomerase Immortalized human Coronary Artery Endothelial cells (TICAE) from the European Collection of Authenticated Cell Cultures (ECACC), and Telomerase Immortalized human Microvascular Endothelial cells (TIME) from the American Type Cell Culture (ATTC), were grown in MesoEndo Cell Growth Medium (Sigma-Aldrich Co., LCC, Diegem, Belgium) at 37°C in a humidified incubator supplemented with 5% CO2.

Techniques: Activity Assay, Multiplex Assay, MANN-WHITNEY, Control

Journal: Frontiers in Pharmacology

Article Title: Connexin43 Hemichannel Targeting With TAT-Gap19 Alleviates Radiation-Induced Endothelial Cell Damage

doi: 10.3389/fphar.2020.00212

Figure Lengend Snippet: Radiation-induced endothelial DNA damage and effect of TAT-Gap19. Gamma H2AX foci, TP53BPI foci and the colocalized Gamma H2AX/TP53BPI foci were assessed in (A) TICAE cells and (B) TIME cells at 1 h after 0.1 and 5 Gy of X-ray exposure with or without applying TAT-Gap19. (C) Representative images showing gamma H2AX foci (green), TP53BPI foci (red) and colocalized gamma H2AX/TP53BPI foci (yellow) in DAPI stained nuclei of TICAE cells at 5 Gy dose. The values represent the average ± SEM of eight biological replicates; statistical significance was analyzed with a non-parametric Mann–Whitney T -test. *Indicates statistically significant differences compared to the respective 0 Gy controls. •Indicates statistically significant differences in the TAT-Gap19 group compared to the corresponding responses in the control group (not treated with TAT-Gap19). */• p < 0.05; **/•• p < 0.01., ***/••• p < 0.001.

Article Snippet: Two human endothelial cell lines: Telomerase Immortalized human Coronary Artery Endothelial cells (TICAE) from the European Collection of Authenticated Cell Cultures (ECACC), and Telomerase Immortalized human Microvascular Endothelial cells (TIME) from the American Type Cell Culture (ATTC), were grown in MesoEndo Cell Growth Medium (Sigma-Aldrich Co., LCC, Diegem, Belgium) at 37°C in a humidified incubator supplemented with 5% CO2.

Techniques: Staining, MANN-WHITNEY, Control

Journal: Frontiers in Pharmacology

Article Title: Connexin43 Hemichannel Targeting With TAT-Gap19 Alleviates Radiation-Induced Endothelial Cell Damage

doi: 10.3389/fphar.2020.00212

Figure Lengend Snippet: Schematic overview of the experimental findings. Blocking Cx43 hemichannel with TAT-Gap19 significantly reduced oxidative stress, inflammation, senescence and cell death in the irradiated coronary artery and microvascular (TICAE and TIME) endothelial cells, and these responses were mainly observed at the high dose (5 Gy).

Article Snippet: Two human endothelial cell lines: Telomerase Immortalized human Coronary Artery Endothelial cells (TICAE) from the European Collection of Authenticated Cell Cultures (ECACC), and Telomerase Immortalized human Microvascular Endothelial cells (TIME) from the American Type Cell Culture (ATTC), were grown in MesoEndo Cell Growth Medium (Sigma-Aldrich Co., LCC, Diegem, Belgium) at 37°C in a humidified incubator supplemented with 5% CO2.

Techniques: Blocking Assay, Irradiation

Journal: Scientific Reports

Article Title: Single and fractionated ionizing radiation induce alterations in endothelial connexin expression and channel function

doi: 10.1038/s41598-019-39317-9

Figure Lengend Snippet: The effect of single and fractionated irradiation on Cx37, Cx40 and Cx43 gene expression. Gene expression of Cx37, Cx40 and Cx43 at 6 h, 24 h, 48 h, 72 h, 7 d or 14 d after a single X-ray exposure (0.1, 0.5 and 5 Gy) in TICAE (left side) and TIME cells (right side) (a,b,c). Gene expression of Cx37, Cx40 and Cx43 at 24 h (d) and 7 d (e) after single or fractionated irradiation in TICAE (left side) and TIME cells (right side). Fractionated irradiation involved three consecutive X-rays doses (0.033 and 1.67 Gy/fraction/day), leading to cumulative doses of 0.1 and 5 Gy. Data were analyzed with a nonparametric Mann-Whitney T-test. Values represent average ± SEM of 5 biological replicates, except for 6 h p.i. where 4 biological replicates were used. ( a – c ) *Indicates for a given time point the statistical difference of gene expression after a dose of single irradiation compared to the respective normalized 0 Gy controls at the same time point. ( d – e ) • Indicates for a given time point the statistical difference of gene expression after a dose of fractionated irradiation compared to the respective normalized 0 Gy controls at the same time point. ( d – e ) *Indicates the statistical difference between fold changes of gene expression after a given radiation dose and a given time of single and fractionated irradiation compared to the respective normalized 0 Gy controls at the same time point. */• p < 0.05; **/•• p < 0.01; ***/••• p < 0.0001. Cx, connexin; TICAE, Telomerase Immortalized human Coronary Artery Endothelial cells; TIME, Telomerase Immortalized human Microvascular Endothelial cells; p.i, post irradiation; h, hours; d, days; SEM, standard error of mean.

Article Snippet: We used two human endothelial cell lines: hTERT telomerase immortalized human coronary artery endothelial cells (TICAE) from the European Collection of Authenticated Cell Cultures (ECACC; HCAECs Cat. No: 300-05), and telomerase immortalized human dermal microvascular endothelial cells (TIME) from the American Type Cell Culture (ATTC).

Techniques: Irradiation, Gene Expression, MANN-WHITNEY