HUVECs (cat. no. BNCC337616), obtained from BeNa Culture Collection, were cultured in DMEM supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin in a humidified incubator with 5% CO2 at 37˚C. HUVECs were treated with different concentrations of Rg1 (1, 5 or 10 µM; Shanghai YaJi Biotechnology Co., Ltd.) for 24 h at 37˚C (18 (link),19 (link)). The AMP-activated protein kinase (AMPK) inhibitor compound C (CC; 5 mM; MilliporeSigma) was used to pretreat HUVECs for 2 h at 37˚C. In addition, HUVECs were stimulated with 100 mg/l ox-LDL (Beijing Solarbio Science & Technology Co., Ltd.) for 48 h at 37˚C to construct an in vitro AS model. To evaluate the effects of Rg1 on ox-LDL-treated HUVECs, the cells were pretreated with Rg1 for 30 min prior to ox-LDL treatment. Untreated cells were regarded as the control group.
Huvecs
Manufactured by Solarbio
Sourced in China, United States
HUVECs are primary human umbilical vein endothelial cells. They are isolated from the human umbilical vein and are used for in vitro studies of endothelial cell biology.
Automatically generated - may contain errors
Lab products found in correlation
Huvecs, by American Type Culture Collection (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Huvecs, by ScienCell (3 mentions)
Ox ldl, by Solarbio (3 mentions)
Rpmi 1640 medium, by Solarbio (2 mentions)
Endothelial cell medium (ecm), by ScienCell (2 mentions)
Heparin, by Selleck Chemicals (1 mentions)
Endothelial cell growth supplement (ecgs), by Thermo Fisher Scientific (1 mentions)
Bleomycin, by Solarbio (1 mentions)
Fin56, by Selleck Chemicals (1 mentions)
F 12k medium, by Thermo Fisher Scientific (1 mentions)
Pkh26, by Umibio (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Huvecs, by iCell Bioscience (1 mentions)
Huvecs, by Thermo Fisher Scientific (1 mentions)
Dexamethasone (dex), by Solarbio (1 mentions)
Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by ScienCell (1 mentions)
Fetal bovine serum (fbs), by ScienCell (1 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (1 mentions)
14 protocols using huvecs
1
Rg1 Modulation of AMPK in Ox-LDL-Induced HUVEC
2
Dexamethasone Treatment of HUVEC Cells
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Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell Corporation (Shanghai, China) and cultured in endothelial cell medium (ECM; ScienCell) containing 5% (v/v) fetal bovine serum (ScienCell), 1% (v/v) endothelial cell growth supplement (ECGS; ScienCell) and 1% penicillin-streptomycin (ScienCell) in a humidified atmosphere of 5% CO2 at 37°C. Transfection of HUVECs was performed using Lipofectamine RNAiMAX Reagent (Invitrogen) in accordance with the manufacturer's protocol. HUVECs treated with 10 μM DEX (Solarbio) and HUVECs without any treatment were termed as the model and control, respectively.
3
In vitro model of atherosclerosis
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Human umbilical vein endothelial cells (HUVECs) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM; Solarbio Science & Technology, Beijing, China), supplemented with 10% fetal bovine serum (FBS; Solarbio Science & Technology) and 1% penicillin-streptomycin (Sangon Biotech, Shanghai, China) in a humidified 5% CO2 incubator at 37 °C. In vitro, the AS model was established by incubation of HUVECs with 100 µg/mL ox-LDL (Sigma-Aldrich, St. Louis, MO, USA) for 24 h. HUVECs were treated with 1 µg/mL LPS (L8880, Solarbio, Beijing, China) for 12 h. HUVECs were cultured in 0.1% DMSO as a control. HUVECs were treated with 1 mM 3-MA (Sigma-Aldrich) for 4 h.
4
Establishing Atherosclerosis Model with HUVECs
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Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA). HUVECs were incubated with RPMI 1640 medium (Solarbio, Beijing, China) containing 10% FBS in an incubator containing 5% CO2 at 37 °C. When the cell growth reached 90% confluence, the medium was discarded, then PBS and trypsin were added respectively for cell washing and digestion, and sub-culturation was performed. To mimic the AS condition in vitro, HUVECs were given 100 mg/L ox-LDL (Yiyuan Biotech, Guangzhou, China) and cultured for 24 h.
5
Lipid Metabonomics in HUVEC Senescence
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Human umbilical vein endothelial cells (HUVECs) were purchased from ATCC (Manassas, VA, USA) and cultured in F-12K Medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS (GIBCO, Grand Island, NY, USA), 0.1 mg/mL heparin (Selleck Chemicals, Houston, TX, USA), and 30 μg/mL Endothelial Cell Growth Supplement (Thermo Fisher) at 37°C in a 5% CO2 atmosphere.
To explore the effects of bleomycin (MedChem Express, NJ, USA) and Astragaloside IV (AS-IV, Solarbio, Beijing, China) on lipid metabonomics in HUVECs, the cells were treated with bleomycin (50 μM) to induce cell senescence; after which, they were coincubated with AS-IV (50 μM) and bleomycin to explore the effect of AS-IV. At the same time, the effect of different doses of bleomycin on LPC levels in HUVECs was detected. The cells were treated with different concentrations of LPC (0.1 μM, 0.25 μM, and 0.4 μM, Merck Millipore, Burlington, MA, USA) to examine the influence of LPC on cell senescence and damage. To further explore how AS-IV functions in the LPC-induced ferroptosis of endothelial cells, the cells were coincubated with LPC (0.4 μM) and AS-IV (50 μM) with or without FIN56 (Selleck, 5 μM).
To explore the effects of bleomycin (MedChem Express, NJ, USA) and Astragaloside IV (AS-IV, Solarbio, Beijing, China) on lipid metabonomics in HUVECs, the cells were treated with bleomycin (50 μM) to induce cell senescence; after which, they were coincubated with AS-IV (50 μM) and bleomycin to explore the effect of AS-IV. At the same time, the effect of different doses of bleomycin on LPC levels in HUVECs was detected. The cells were treated with different concentrations of LPC (0.1 μM, 0.25 μM, and 0.4 μM, Merck Millipore, Burlington, MA, USA) to examine the influence of LPC on cell senescence and damage. To further explore how AS-IV functions in the LPC-induced ferroptosis of endothelial cells, the cells were coincubated with LPC (0.4 μM) and AS-IV (50 μM) with or without FIN56 (Selleck, 5 μM).
6
Endothelial Cell Response to Uremic Factors
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HUVECs were originally obtained from Sciencell Research Laboratories, and cultured in endothelial cell medium (Sciencell, California, USA) containing 5% fetal bovine serum, 1% penicillin-streptomycin solution and endothelial cell growth supplement in a 5% CO2 incubator. HUVECs at passages 5–8 were used for the in vitro studies. To examined the effect of uremic serum on endothelial cells, sera from patients with CKD or controls were pooled, and diluted to 20% by using ECM culture media, then endothelial cells were incubated for 48 h. IS and Hcy (Sigma-Aldrich, St. Louis, MO), IAA and HA (Solarbio, Beijing, China) were diluted in ECM media and used to treated HUVECs. The cells were verified by short tandem repeat profiling and tested negative for mycoplasma contamination.
7
Exosome Labeling and Internalization in DPSCs and HUVECs
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A total of 500 μg of M1-Exos and M2-Exos were labeled with the lipophilic dye PKH-26 (Umibio, Shanghai, China), respectively, followed by ultracentrifugation to remove excess dye. Then, the PKH-26-labeled exosomes were added to DPSCs or commercially obtained HUVECs (iCell, Shanghai, China) and incubated for 24 h. Afterward, the co-incubated cells were washed three times with PBS and subsequently fixed with PFA for 20 min. Next, fluorescent phalloidin (Abbkine, Wuhan, China) was employed to label the actin filament of the cytoskeleton in DPSCs and HUVECs for 30 min, and DAPI (Solarbio, Beijing, China) was utilized for staining cell nuclei for 5 min. Finally, the cells were rinsed three times with PBS and photographed under a laser confocal microscope.
8
Oxidized LDL Exposure in HUVECs
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Human umbilical vein endothelial cells (HUVECs) acquired from American Type Culture Collection (ATCC; Manassas, VA, USA) were cultivated in Dulbecco's modified Eagle's medium (DMEM) plus 10% FBS (Gibco, Carlsbad, CA, USA) and 1% antibiotics (Gibco) under 37 °C with 5% CO2. The ox-LDL group was generated by exposing HUVECs to 100 μg/mL ox-LDL (Solarbio) for 24 h [18 ,19 ] to study cellular responses.
9
Cultivation and Characterization of Cell Lines
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The HNSCC cell line Fadu, established a hypopharyngeal tumor from an Indian individual, was purchased from the Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China). Human Thp-1 monocytes, derived from peripheral blood of a 1-year-old boy, were purchased from the Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China). The Cal-27 cell line, established from a primary tongue cancer, was donated by the Institute of Stomatology, Nanjing Medical University, and purchased from American Type Culture Collection (ATCC; Manassas, USA). The human umbilical vein endothelial cell line (HUVECs) was donated by Dr. Hong Ji of Fudan University and purchased from ATCC (Manassas, USA). Fadu and Cal-27 cells were cultured in DMEM, while HUVECs were maintained in M199 medium (Solarbio, Beijing, China). Thp-1 cells were cultured in RPMI-1640 medium (Solarbio, Beijing, China). All cultures were supplemented with 10% fetal bovine serum (Gibco, Rockville, MD) and 1% penicillin and streptomycin (Gibco), and maintained at 37°C in 5% CO2. DHA was purchased from Tokyo Chemical Industry (Tokyo, Japan).
10
Evaluating Cell Viability on Biomaterial Films
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For the long-term test, the subject was asked to behave the same as daily life, including work, exercise and taking shower regularly. The sEMG recording was done every noon to monitor performances of electrodes. For cell viability tests, PPA, LM, PDMS, PEDOT-PVA and TPP films were sterilized under UV light overnight. Especially, the TPP film was treated with desterilized FeCl3 solution (10 mg/mL) for 48 h to stabilize the film on a slide glass. HUVECs (ATCC, USA) were seeded at 105 mL−1 on the film and cultured in Dulbecco’s modified eagle medium (DMEM, Gibco, USA) supplemented with 10% foetal bovine serum (5% CO2, 37 oC). After 5 days, HUVECs were stained with live/dead kit: Calcein-AM (green, live cells, Solarbio, China), PI (red, dead cells, Solarbio, China), and DAPI (blue, nucleus, Sigma, USA). The confocal microscope (ZEISS LSM 980, Germany) was used to take fluorescent images.
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