human lung microvascular endothelial cells hlmvecs  (Cell Applications Inc)

Journal: Scientific Reports

Article Title: A next-generation system for smoke inhalation integrated with a breathing lung-on-chip to model human lung responses to cigarette exposure

doi: 10.1038/s41598-025-00438-z

Figure Lengend Snippet: Apical surfactant addition demonstrates potential protective effects against CS exposure. ( A ) Overview of cells used in the triple culture “TC THP1” (AXiAECs/hLMVECs/THP1), culture conditions (dynamic; ALI + Str) and treatment (Surfactant pre-treated followed by CS exposure) performed. ( B ) Representative immunofluorescence staining of CTRL ALI + Str (− Surfactant) cells (top row; scale bar 50 µm), CS exposed ALI + Str (-Surfactant) cells (second row; scale bar 50 µm), CTRL ALI + Str (+ Surfactant) cells (third row; scale bar 100 µm), CS exposed ALI + Str (+ Surfactant) cells (fourth row; scale bar 100 µm). Cells were stained for Zonula Occludens 1 (ZO-1, in green) and nuclei (Hoechst, in blue). ( C ) IL-8 concentration (pg/ml) measured from apical supernatants collected from the CTRL and CS exposed wells pre-treated with and without surfactant (+ Surf or -Surf) via ELISA assay (N = 2; n = 6/condition). ( D ) mRNA was harvested at 48 h time-point with and without surfactant (+ Surf or -Surf) post-exposure to CS. qRT-PCR study for inflammation (Interleukin 6, IL6; Interleukin 8, IL8; Tumor Necrosis Factor α, TNFα) and oxidative stress-related (Heme Oxygenase 1, HO-1; Human MutT Homolog 1, MTH-1) genes were conducted in all -Surf and + Surf CS exposed and CTRL samples (N = 2, n = 3). Data are shown as mean ± SEM.

Article Snippet: The AX iAECs were cultured according to the manufacturer’s instructions, as previously described and characterized by Sengupta et al. . Human Lung Microvascular Endothelial Cells (hLMVECs) purchased from PromoCell were expanded in flasks using AX Endothelial Medium. hLMVECs were first seeded on the basolateral side of each membrane in the AX12 at a density of 106,000cells/cm 2 , followed by a 2-h incubation, and then AX iAECs were seeded apically at a density of 409,000 cells/cm 2 on day 0 for all co-culture experiments.

Techniques: Immunofluorescence, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: A next-generation system for smoke inhalation integrated with a breathing lung-on-chip to model human lung responses to cigarette exposure

doi: 10.1038/s41598-025-00438-z

Figure Lengend Snippet: Differential effects of CS exposure on various co-culture dynamic models. ( A ) Overview of cells used in the dual culture “DC” ( AX iAECs/hLMVECs), culture conditions (dynamic; ALI + Str) and treatment (CS exposure) performed. ( B ) Overview of cells used in the triple culture “TC pBDMs” ( AX iAECs/hLMVECs/pBDMs) and “TC THP1” ( AX iAECs/hLMVECs/THP1), culture conditions (dynamic; ALI + Str) and treatment (CS exposure) performed. ( C ) TER (Ohm cm 2 ) measured in “ALI + Str” conditions pre-exposure at 0 h and 4 h, 24 h and 48 h post-exposure to CS in DC (N = 3; n = 3/time-point), in TC pBDMs (N = 2; n = 3/time-point) and TC THP1 (N = 3; n = 3/time-point). Data are shown as violin plots, medians are indicated by black dotted lines. ( D ) The levels of IL-8 (pg/ml) in the supernatants collected from the cells were measured by ELISA in all the co-culture models. ( E ) Cytotoxicity was calculated from LDH release at 48 h time-point after CS exposure (N = 2; n = 3/time-point for all models). Data are shown as mean ± SEM.

Article Snippet: The AX iAECs were cultured according to the manufacturer’s instructions, as previously described and characterized by Sengupta et al. . Human Lung Microvascular Endothelial Cells (hLMVECs) purchased from PromoCell were expanded in flasks using AX Endothelial Medium. hLMVECs were first seeded on the basolateral side of each membrane in the AX12 at a density of 106,000cells/cm 2 , followed by a 2-h incubation, and then AX iAECs were seeded apically at a density of 409,000 cells/cm 2 on day 0 for all co-culture experiments.

Techniques: Co-Culture Assay, Enzyme-linked Immunosorbent Assay

Journal: Redox Biology

Article Title: Human lung microvascular endothelial cell protein modification by 2-chlorohexadecanoic acid: RhoA mediates 2-chlorohexadecanoic acid-elicited endothelial activation

doi: 10.1016/j.redox.2025.103596

Figure Lengend Snippet: 2-ClHyA modified HLMVEC proteins . (A) Venn diagram of the 2-ClHyA- and HyA-modified proteins in HLMVECs. (B) Protein class distributions of the 11 proteins identified exclusively in the 2-ClHyA sample group, analyzed using PANTHER classification system. (C) Protein class distributions of the 194 proteins identified in both the 2-ClHyA and HyA sample groups, analyzed using PANTHER.

Article Snippet: Human lung microvascular endothelial cells (HLMVEC) (PromoCell, C‐12281, Heidelberg, Germany) were cultured at 37 °C in EGM‐2MV medium (Sigma, cat. CC‐3202), in a humidified atmosphere with 95 % air/5 % CO 2 (v/v).

Techniques: Modification

Journal: Redox Biology

Article Title: Human lung microvascular endothelial cell protein modification by 2-chlorohexadecanoic acid: RhoA mediates 2-chlorohexadecanoic acid-elicited endothelial activation

doi: 10.1016/j.redox.2025.103596

Figure Lengend Snippet: Bioinformatic characterization of HLMVEC proteins modified by 2-ClHyA and HyA: Enriched Gene Ontology (GO) cellular components are shown, and were the only GO categories enriched in the 2-ClHyA modified proteome. Over-representation analysis was performed with the A.GO. TOOL using Bonferroni p < 0.05 for computing FDR with a p value cutoff of 0.01. Over-representation was compared to the HLMVEC proteome as a background. S value is a combination of -log p value and effect size. Effect size represents the difference in proportions of the foreground and background proteomes.

Article Snippet: Human lung microvascular endothelial cells (HLMVEC) (PromoCell, C‐12281, Heidelberg, Germany) were cultured at 37 °C in EGM‐2MV medium (Sigma, cat. CC‐3202), in a humidified atmosphere with 95 % air/5 % CO 2 (v/v).

Techniques: Modification

Journal: Redox Biology

Article Title: Human lung microvascular endothelial cell protein modification by 2-chlorohexadecanoic acid: RhoA mediates 2-chlorohexadecanoic acid-elicited endothelial activation

doi: 10.1016/j.redox.2025.103596

Figure Lengend Snippet: Protein-protein association network of the proteins modified exclusively by 2-ClHyA in HLMVEC. A) Association network of all proteins modified by 2-ClHyA (205 proteins). Red nodes indicate proteins identified in the cell-cell junction family of the Subcellular location group. Blue nodes indicate proteins identified in the actin filament-based process family of the Biological Process (GO) group. Green nodes indicate proteins identified in the proteasomal protein catabolic process family of the Biological Process (GO) group. Yellow nodes indicate proteins identified in the generation of precursor metabolites and energy family of the Biological Process (GO) group. Network generated using STRING, with confidence set to medium. B) Association network of proteins exclusively modified by 2-ClHyA (11 proteins). Blue nodes indicate proteins identified in the tight junction family of the KEGG Pathways. Network generated using STRING, with confidence set to medium. RhoA is highlighted in a red rectangle in STRING networks in both A and B. C) List of the 11 proteins exclusively modified by 2-ClHyA. PSM denotes peptide spectrum map.

Article Snippet: Human lung microvascular endothelial cells (HLMVEC) (PromoCell, C‐12281, Heidelberg, Germany) were cultured at 37 °C in EGM‐2MV medium (Sigma, cat. CC‐3202), in a humidified atmosphere with 95 % air/5 % CO 2 (v/v).

Techniques: Modification, Generated

Journal: Redox Biology

Article Title: Human lung microvascular endothelial cell protein modification by 2-chlorohexadecanoic acid: RhoA mediates 2-chlorohexadecanoic acid-elicited endothelial activation

doi: 10.1016/j.redox.2025.103596

Figure Lengend Snippet: RhoA mediated 2-ClHA-elicited HLMVEC barrier leak. A) HLMVECs were grown on ECIS arrays to confluency and then were either pre-treated with C3 or vehicle (arrow 1) 4h prior to lipid addition, Rhosin or vehicle (arrow 2) 2h prior to lipid addition. Treatment with 2-ClHA (10 μM) or vehicle is indicated by arrow 3. Resistance is normalized to levels measured 30 min prior to starting pretreatment. Data are mean±S.E.M.; n = 20. B) Individual data points following 6h of treatment conditions in panel A are shown as well as other conditions including Rhosin and C3 treatment without lipid treatment as well as indicated concentrations of either 2-ClHA or HA. Average change in normalized resistance at 6h following lipid treatment, data are mean±S.E.M.; n = 20 independent experiments. C) RhoA activity in HLMVEC treated with either BSA (vehicle), 2-ClHA, or HA (10 μM) was measured as described in “Materials and Methods”. HLMVECs were pretreated with C3 (1 μg/ml) or Rhosin (30 μM) 4h or 2h, respectively prior to lipid treatments. Data are mean±S.E.M.; n = 8–9 independent experiments. D) Ang-2 level in HLMVEC cell culture media was determined in cells treated with either BSA (vehicle), 2-ClHA, or HA (10 μM) was measured as described in “Materials and Methods”. HLMVECs were pretreated with C3 (1 μg/ml) or Rhosin (30 μM) 4h or 2h, respectively prior to lipid treatments. Data are mean±S.E.M.; n = 5 independent experiments. ∗, ∗∗∗, and ∗∗∗∗ indicate p < 0.05, p < 0.001, and p < 0.0001 for indicated comparisons.

Article Snippet: Human lung microvascular endothelial cells (HLMVEC) (PromoCell, C‐12281, Heidelberg, Germany) were cultured at 37 °C in EGM‐2MV medium (Sigma, cat. CC‐3202), in a humidified atmosphere with 95 % air/5 % CO 2 (v/v).

Techniques: Activity Assay, Cell Culture