Journal: The American Journal of Tropical Medicine and Hygiene

Article Title: Dengue Virus Infection of Primary Human Smooth Muscle Cells

doi: 10.4269/ajtmh.18-0175

Figure Lengend Snippet: Human umbilical artery smooth muscle cells (HUASMC) are permissive to dengue virus (DENV) infection. Virion production and release was quantified by plaque assays of culture supernatants at different time points post-infection (p.i.) of HUASMC, human umbilical vein endothelial cells (HUVEC), and LLC-MK2 (macaque kidney cells) infected with each of the four DENV serotypes (multiplicity of infection:1). (A) Infection kinetics of HUASMC cells (24–72 hours p.i.) with DENV 1–4 measured by plaque assays (plaque-forming units [PFU]/mL). (B) DENV 1–4 titers in culture supernatants of HUASMC, HUVEC, and LLC-MK2 cells at 72 hours p.i. Data are expressed as the mean ± standard deviation of three independent experiments. ***P < 0.001 compared with its HUVEC and LLC-MK2 cells counterparts. (D) l. = assay detection limit.

Article Snippet: Human umbilical artery smooth muscle cells and HUVEC were purchased and maintained in smooth muscle cell growth medium and endothelial cell growth medium, respectively, according to the manufacturer’s instructions (Cell Applications, San Diego, CA).

Techniques: Virus, Infection, Standard Deviation

Journal: The American Journal of Tropical Medicine and Hygiene

Article Title: Dengue Virus Infection of Primary Human Smooth Muscle Cells

doi: 10.4269/ajtmh.18-0175

Figure Lengend Snippet: Human umbilical artery smooth muscle cells (HUASMC) release dengue virus (DENV) genomes on infection. Dengue virus genomic RNA was quantified by real-time qRT-PCR from cell culture supernatants of HUASMC, human umbilical vein endothelial cells (HUVEC), and LLC-MK2 (macaque kidney cells) infected with DENV (multiplicity of infection:1). (A) Dengue virus 1–4 infection kinetics (24–72 hours post-infection [p.i.]) of HUASMC cells measured by real-time genomic qRT-PCR (copies/mL). (B) Genome copies present in culture supernatants of HUASMC, HUVEC, and LLC-MK2 cells at 72 hours p.i. with the four DENV serotypes. (C) Calculated genome-to-plaque-forming unit (PFU) ratios of HUASMC, HUVEC, and LLC-MK2 cells supernatants at 72 hours p.i. with each DENV serotype. Data are expressed as the mean ± standard deviation of three independent experiments. *P < 0.05, **P < 0.005, and ***P < 0.001 calculated to its HUASMC counterpart.

Article Snippet: Human umbilical artery smooth muscle cells and HUVEC were purchased and maintained in smooth muscle cell growth medium and endothelial cell growth medium, respectively, according to the manufacturer’s instructions (Cell Applications, San Diego, CA).

Techniques: Virus, Infection, Quantitative RT-PCR, Cell Culture, Standard Deviation

Journal: The American Journal of Tropical Medicine and Hygiene

Article Title: Dengue Virus Infection of Primary Human Smooth Muscle Cells

doi: 10.4269/ajtmh.18-0175

Figure Lengend Snippet: Dengue virus (DENV) antigens are detected in human umbilical artery smooth muscle cells (HUASMC). Epifluorescence images of immunostained cells with an anti-DENV 1-4 envelope protein-specific monoclonal antibody (green), and a cytoplasmic (red) and nuclei (blue) counterstains. The images were captured at 72 hours post-infection (p.i.) with each DENV serotype at a multiplicity of infection (MOI) of 1. (A) Representative image of mock-infected HUASMC cells at ×100 magnification (scale bar = 50 μm). (B and C) Representative images of DENV-infected HUASMC (arrows) at ×400 (scale bar = 20 μm) and ×100 (scale bar = 50 μm) magnification, respectively. (D) Image analysis for quantifying infected cells (green outline) vs total cell nuclei (white outlines) with the software CellProfiler 2.0. (E) Percentages of infected cells in HUASMC, human umbilical vein endothelial cells (HUVEC), and LLC-MK2 (macaque kidney cells) cell lines. (F) Comparison of DENV labeling by immunostaining with an anti-DENV 1-4 envelope protein-specific monoclonal antibody (green) and an anti-NS3 polyclonal antibody (red) in HUASMC cells at 72 hours p.i. with DENV-2 and DENV-3 at a MOI of 1. Magnification of ×400 (scale bar = 20 μm). Data are expressed as mean ± standard deviation of three independent experiments. **P < 0.005, ***P < 0.001 calculated to its HUASMC cells counterpart. This figure appears in color at www.ajtmh.org.

Article Snippet: Human umbilical artery smooth muscle cells and HUVEC were purchased and maintained in smooth muscle cell growth medium and endothelial cell growth medium, respectively, according to the manufacturer’s instructions (Cell Applications, San Diego, CA).

Techniques: Virus, Infection, Software, Comparison, Labeling, Immunostaining, Standard Deviation

Journal:

Article Title: Development of Composite Porous Scaffolds Based on Collagen and Biodegradable Poly(ester urethane)urea

doi:

Figure Lengend Snippet: Growth curves for scaffolds made from PEUU (C0/ 1080) and PEUU with 10% collagen (C10/1080) seeded with umbilical artery smooth muscle cells and cultured for up to 28 days. The relative cell number is based upon MTT absorption and reflects mitochondrial activity.

Article Snippet: Scaffolds were cut and sterilized by immersion in 70% ethanol for 6 h, followed by rinsing with PBS and exposure to the ultraviolet light source in a laminar flow hood (Class II A/B3 Biological Safety Cabinet) for 2 h. Scaffolds were placed in a centrifuge tube with human umbilical artery smooth muscle cells (UASMCs, Bio Whittaker) at a density of 1 × 10 6 cells/ml in medium (SmGM-2 with 10% fetal bovine serum, BioWhittaker).

Techniques: Cell Culture, Activity Assay

Journal:

Article Title: Development of Composite Porous Scaffolds Based on Collagen and Biodegradable Poly(ester urethane)urea

doi:

Figure Lengend Snippet: Electron micrographs of PEUU and PEUU/collagen (90:10) scaffold surfaces after 28 days of culture with umbilical artery smooth muscle cells.

Article Snippet: Scaffolds were cut and sterilized by immersion in 70% ethanol for 6 h, followed by rinsing with PBS and exposure to the ultraviolet light source in a laminar flow hood (Class II A/B3 Biological Safety Cabinet) for 2 h. Scaffolds were placed in a centrifuge tube with human umbilical artery smooth muscle cells (UASMCs, Bio Whittaker) at a density of 1 × 10 6 cells/ml in medium (SmGM-2 with 10% fetal bovine serum, BioWhittaker).

Techniques:

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: cGMP accumulation in co‐cultures of human primary vascular smooth muscle cells following addition of serelaxin to endothelium. HUAEC , HUVEC or HCAEC were co‐cultured with (A) HUASMC or (B) HUVSMC (all n = 5), and the ECs were treated with serelaxin for 30 min. Serelaxin addition to HUAEC did not cause cGMP accumulation in HUAEC (▲) (C) HUASMC (□) or (D) HUVSMC (◯) co‐cultured with HUAEC, whereas direct stimulation of either (C) HUASMC (n = 5) or (D) HUVSMC with serelaxin caused a concentration‐dependent increase in cGMP accumulation (dashed lines). In contrast, serelaxin addition to HUVEC concentration‐dependently increased cGMP accumulation not only in HUVEC (■) but also in (E) HUASMC (□) or (F) HUVSMC (◯) co‐cultured with HUVEC with the responses in smooth muscle cells being greater or in the case of HUVSMC much greater than cGMP responses to direct stimulation of (E) HUASMC or (F) HUVSMC (dashed lines). A similar pattern of cGMP accumulation was observed with (G, H) HCAEC (●) and (G) HUASMC (□) or (H) HUVSMC (◯) co‐cultured with HCAEC.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Cell Culture, Concentration Assay

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: Serelaxin‐mediated cGMP accumulation in monocultures of human primary vascular cells (all n = 5). Serelaxin (30 nM, 30 min) increased cGMP accumulation in (A) HUVEC, (B) HCAEC, (C) HUASMC and (D) HUVSMC. Pre‐incubation with l‐NOARG (30 μM, 30 min) or ODQ (1 μM, 30 min) almost abolished serelaxin‐mediated (30 nM, 30 min) cGMP accumulation in all cell types. Pre‐treatment with indomethacin (30 μM, 30 min) significantly inhibited serelaxin‐mediated (30 nM, 30 min) cGMP accumulation in (B) HCAEC but had no effect in (A) HUVEC, (C) HUASMC or (D) HUVSMC. *P < 0.05, **P < 0.02, ***P < 0.005; significantly different from serelaxin alone; one‐way anova with Dunnett's post hoc test.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Incubation

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: Serelaxin‐mediated cGMP accumulation in human primary vascular smooth muscle cells co‐cultured with HUVEC or (A) HCAEC (all n = 6 except where otherwise indicated). Stimulation of HUVEC or HCAEC with serelaxin (30 nM, 30 min) increased cGMP accumulation not only in (B) HUVEC and (C) HCAEC but also in co‐cultures of (D, F) HUASMC or (E, G) HUVSMC. Pre‐incubation of HUVEC or HCAEC with l‐NOARG (30 μM, 30 min) before addition of serelaxin (30 nM, 30 min) significantly inhibited cGMP accumulation not only in HUVEC and (C) HCAEC but also in (D, F) HUASMC and (E, G) HUVSMC. Pre‐incubation of HUVEC with indomethacin (30 μM, 30 min) did not affect serelaxin‐mediated (30 nM, 30 min) cGMP accumulation in (B) HUVEC or in co‐incubated (D) HUASMC or (E) HUVSMC (n = 5). Pre‐incubation of HCAEC with indomethacin (30 μM, 30 min) had no significant effect on serelaxin‐mediated (30 nM, 30 min) cGMP accumulation in (C) HCAEC but produced marked and significant reductions in cGMP accumulation in co‐incubated (F) HUASMC or (G) HUVSMC (n = 5). Pre‐treatment of HUASMC or HUVSMC with ODQ (1 μM, 30 min) had no significant effect on serelaxin‐mediated (30 nM, 30 min) cGMP accumulation in (I) HUVEC or (J) HCAEC but reduced or abolished cGMP accumulation in (K, M) HUASMC or (L, N) HUVSMC (n = 5). *P < 0.05, **P < 0.02, ***P < 0.005 significantly different from serelaxin alone; one‐way anova with Dunnett's post hoc test.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Cell Culture, Incubation, Produced

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: cAMP accumulation in co‐cultures of human primary vascular smooth muscle cells following addition of serelaxin to endothelium (all n = 5). HUAEC, HUVEC or HCAEC were co‐cultured with (A) HUASMC or (B) HUVSMC, and the endothelial cells were treated with serelaxin for 30 min. Serelaxin added to HUAEC did not cause cAMP accumulation either in (C, D) HUAEC (▲), (C) HUASMC (□) or (D) HUVSMC (◯), whereas direct stimulation of (C) HUASMC or (D) HUVSMC with serelaxin caused a concentration‐dependent increase in cAMP accumulation (dashed lines). Although direct addition of serelaxin to HUVEC concentration‐dependently increased cAMP accumulation in (E, F) HUVEC (■), there was no significant effect on cAMP accumulation in (E) HUASMC (□) or (F) HUVSMC (◯). Direct addition of serelaxin to (E) HUASMC or (F) HUVSMC stimulated cAMP accumulation (dashed lines). Serelaxin concentration‐dependently increased cAMP accumulation in (G, H) HCAEC (●) but also caused a robust concentration‐dependent increase in cAMP accumulation in both (G) HUASMC (□) and (H) HUVSMC (◯).

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Cell Culture, Concentration Assay

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: Serelaxin‐mediated cAMP accumulation in monocultures of human primary vascular cells (all n = 5). Serelaxin (30 nM, 30 min) increased cAMP accumulation in (A) HUVEC, (B) HCAEC, (C) HUASMC and (D) HUVSMC that was not significantly altered by pre‐incubation with l‐NOARG (30 μM, 30 min) or ODQ (1 μM, 30 min). Pre‐treatment with indomethacin (30 μM, 30 min) significantly inhibited serelaxin‐mediated (30 nM, 30 min) cAMP accumulation in (B) HCAEC but not in (A) HUVEC, (C) HUASMC or (D) HUVSMC. *P < 0.05; significantly different from serelaxin alone; one‐way anova with Dunnett's post hoc test.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Incubation

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: Serelaxin‐mediated cAMP accumulation in human primary vascular smooth muscle cells co‐cultured with HCAEC (A, E; all n = 5). Stimulation of HCAEC with serelaxin (30 nM, 30 min) increased cAMP accumulation not only in (B) HCAEC but also in co‐cultures of (C) HUASMC or (D) HUVSMC. Pre‐incubation of HCAEC with l‐NOARG (30 μM, 30 min) before addition of serelaxin (30 nM, 30 min) had no significant effect on cAMP accumulation in (B) HCAEC, (C) HUASMC or (D) HUVSMC. However, pre‐incubation of HCAEC with indomethacin (30 μM, 30 min) significantly inhibited serelaxin‐mediated (30 nM, 30 min) cAMP accumulation in (B) HCAEC and abolished cAMP accumulation in (C) HUASMC or (D) HUVSMC. Pre‐treatment of HUASMC or HUVSMC with ODQ (1 μM, 30 min) had no significant effect on serelaxin‐mediated (30 nM, 30 min) cAMP accumulation in (F) HCAEC, (G) HUASMC or (H) HUVSMC. *P < 0.05; significantly different from serelaxin alone; one‐way anova with Dunnett's post hoc test.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Cell Culture, Incubation

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: Signal transduction mechanisms activated by serelaxin in co‐cultures of human primary vascular cells. Activation of RXFP1 by serelaxin in HUVEC and HCAEC stimulates NO production and activates sGC and AC to produce cGMP and cAMP respectively. Endothelial NO also diffuses from the endothelial cells across the ThinCert membranes and activates sGC in both the arterial and venous smooth muscle cells. Additionally in HCAEC (blue lines) but not HUVEC, serelaxin stimulates prostanoid production that produces cAMP accumulation in both arterial and smooth muscle cells.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Transduction, Activation Assay

Journal: Bioactive Materials

Article Title: A “built-up” composite film with synergistic functionalities on Mg–2Zn–1Mn bioresorbable stents improves corrosion control effects and biocompatibility

doi: 10.1016/j.bioactmat.2023.02.004

Figure Lengend Snippet: (A) Fluorescent staining of HUVECs cultured for 1 day, 3 days, and 5 days. (B) HUVECs' viability after 1 day, 3 days, and 5 days of incubation is calculated from CCK-8 tests. (C) Fluorescent staining of HUASMCs cultured for 1 day, 3 days, and 5 days. (D) HUASMCs' viability after 1 day, 3 days, and 5 days of incubation is calculated from CCK-8 tests. Data are presented as mean ± SD (n = 4), *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: In the cell culture experiment, human umbilical vein endothelial cells (HUVECs) and human umbilical artery smooth muscle cells (HUASMCs) purchased from Guangzhou Geneo Biotech were first subcultured and then seeded in 24-well plates at a density of 1.5 × 10 4 and 2 × 10 4 cells/ml, respectively.

Techniques: Staining, Cell Culture, Incubation, CCK-8 Assay