Human umbilical vein endothelial cells (HUVECs) were obtained from Cell Applications (San Diego, CA). Tube formation assays were performed according to previously published protocols79 (link). Briefly, HUVECs were seeded at 2 × 105 cells per 75 cm2 in T75 culture flasks and expanded in Human EC Growth Medium (Cell Applications) until 75% confluence was reached. After washing off growth media, cells were serum starved one day before the assay by incubation in Human EC Basal Medium (Cell Applications) for 24 h. Calcein AM (green; ThermoFisher) was added to cultures at a concentration of 2 µg/ml to stain cells and assess cell viability. Cells were incubated with Calcein AM in the dark at 37 °C and 5% CO2 for 30 min, then the stain was washed off using PBS. Assays were performed in 24-well plates coated with Geltrex LDEV (lactose dehydrogenase elevating virus)-Free Reduced Growth Factor Basement Membrane Matrix (ThermoFisher) at 75 µl/cm2 according to the manufacturer’s recommendations. Cells were plated at a concentration of 3.5 × 104 in 200 µl Human EC Basal Medium (Cell Applications) per well. For co-culture assays, DCs were stained using CellTrace Calcein Red-Orange (ThermoFisher) and added to the EC cultures at a 1:1 ratio. Assays were run for 12 h and then fixed using 4% paraformaldehyde and imaged using a Zeiss LSM 880 confocal laser scanning microscope.
Huvecs
Manufactured by Cell Applications
Sourced in United States
HUVECs are primary human umbilical vein endothelial cells. They are a common in vitro model for studying endothelial cell biology and function.
Automatically generated - may contain errors
Lab products found in correlation
Endothelial cell growth medium, by Cell Applications (5 mentions)
Huvecs, by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Huvecs, by Merck Group (4 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Egm 2mv, by Lonza (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Penicillin and streptomycin, by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Egm bulletkit, by Lonza (2 mentions)
Fibroblast growth kit low serum, by American Type Culture Collection (2 mentions)
Huaecs, by Cell Applications (2 mentions)
Phosphate buffered saline (pbs), by Merck Group (2 mentions)
C 12231, by PromoCell (2 mentions)
C 12271, by Lonza (2 mentions)
Hcoaecs, by Lonza (2 mentions)
Panx1 silencer, by Thermo Fisher Scientific (2 mentions)
Gja1 silencer, by Thermo Fisher Scientific (2 mentions)
Haoec, by PromoCell (2 mentions)
55 protocols using huvecs
1
HUVEC Tube Formation Assay Protocol
2
Culturing Human and Mouse Endothelial Cells
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Human Umbilical Vein endothelial cells (HUVECs) were purchased and cultured in an all-in-one complete endothelial growth medium that contains 5 mM Glucose (Cat No.: 211-500) from Cell Applications Inc. (San Diego, CA, USA). HUVECs were purchased from Cell Applications and cultured in HUVEC growth medium [8 (link)]. C57BL/6 Mouse Primary Skeletal Muscle Microvascular Endothelial Cells (Cat No.: C57-6220) were purchased and cultured in Complete Mouse Endothelial Cell Medium (Cat: M1168) from Cell Biologics (Chicago, IL, USA).
3
HUVEC Hypoxia Response Protocol
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Human umbilical vein ECs (HUVECs) were purchased from Cell Applications and used between passages 3 and 6. HUVECs were cultured in All-In-One Endothelial Growth Medium (Cell Applications, catalog no. 211-500). Upon reaching confluence, media were changed to endothelial starvation media (Cell Applications, catalog no. 209-250) and cells were incubated in a 2% oxygen chamber (BioSpherix) for 0, 24, or 48 hours. HUVECs were directly lysed in TRIzol, and total RNA isolation then proceeded using the PureLink total RNA purification system with the on-column DNase protocol according to the manufacturer’s instructions. RNA concentration and purity were determined with a NanoDrop spectrophotometer in duplicate.
4
Endothelial Cell Culture and Passage
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HCAECs and HUVECs were purchased from Cell Applications and cultured in Meso Endo Cell Growth media (212–500) and Endothelial Cell media (211–500), respectively. These cells were cultured in sterile vessels precoated with 0.001% fibronectin (Sigma-Aldrich, F0895) in HBSS (Sigma-Aldrich, H6648) for at least an hour. For passaging, the cells were collected using Trypsin-EDTA (Sigma-Aldrich, T4174) and neutralized with soybean trypsin inhibitor (Invitrogen, 17075–029).
5
HUVEC Apoptosis and Vesicle Generation
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Human umbilical vein endothelial cells (HUVECs) were purchased from Cell Applications, cultured in Medium 200 + LSGS (Gibco, Waltham, MA, USA) on a gelatin-coated surface, and Passage 4 cells were used in experiments. Endothelial cells were exposed to RPMI serum-free medium (Gibco) or Medium 200 + LSGS (depleted of vesicles) for 4 h to produce apoptotic-cell- or healthy-cell-conditioned medium respectively as described previously [13 (link), 15 (link), 18 (link), 19 (link)]. To generate fluorescence-emitting vesicles, HUVECs were stained using CellTracker Orange-CMRA dye or SYTO RNASelect Green Fluorescent cell stain (Molecular Probes, Waltham, MA, USA) for 15 min before treatment. Cytochalasin D, monodansylcadaverine, methyl-β-cyclodextrin (MβCD), and 5-(N-ethyl-N-isopropyl)amiloride (EIPA) were purchased from Sigma (Burlington, MA, USA). FITC-Transferrin was obtained from Thermo Fisher (Waltham, MA, USA), and annexin V was obtained from BioLegend (San Diego, CA, USA).
6
Quercetin Ameliorates HFD-Induced Cardiac Dysfunction
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For in vivo and ex vivo studies (Figure 1 ), four groups of mice were studied (n = 6 − 8 each): control+vehicle (CV), control+quercetin (CQ), HFD + vehicle (HV), and HFD + quercetin (HQ). LV cardiac function was evaluated in vivo using high-field magnetic resonance imaging, whereas intramyocardial fat deposition, microvascular density, oxidative stress, senescence, and histology were estimated ex vivo. In vitro, human umbilical vein endothelial cells (HUVECs, Cell Applications, San Diego, CA; Cat.# 200 K-05f) were treated with oxidized low-density lipoprotein (ox-LDL, Thermo Fisher Scientific, Grand Island, NY; Cat.# L34357) to impair capillary generation, and tube formation was assessed to estimate the angiogenic activity of Q.
7
Co-culture of bMSCs and HUVECs for LPS-stimulation
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Commercialized primary human bMSCs (HUXMA-01001, Cyagen, China) were cultured in a growth medium for human MSCs (HUXMA-90011, Cyagen, China), and cells at passages 3–6 were used for experiments. Human umbilical vein endothelial cell lines (HUVECs, Cell Applications, USA) were cultured in DMEM/F12 medium (Gibco, USA) in an environment of the humidified atmosphere at 37 °C with 5% CO2. All culture mediums contained 10% EV-free FBS (Exo-FBS-50a-1, BI, USA) with 1% penicillin–streptomycin (Gibco, USA). For co-culture experiments, bMSCs were seeded on Transwell inserts in six-well plates (3450, Corning, USA) and HUVECs were seeded in six-well plates. When HUVECs attained 80% confluency, bMSCs and HUVECs were co-cultured in a refreshing medium. HUVECs were stimulated with LPS (1 μg/mL) for 6 h and then were collected for related experiments.
8
HUVEC Cultivation for Research
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Primary cultured human umbilical vein endothelial cells (HUVECs) were purchased from Cell Applications, INC. The HUVECs were cultured in Medium 200 with 1X low-serum growth supplement (LSGS) in 10 mm dishes pre-coated with 0.1% gelatin as previously described28 (link). HUVECs at passages 3–6 were used for this study.
9
Cell Culture of Haploid Leukemia and Endothelial Cells
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Myelogenous leukemia-derived, near-haploid human HAP1 cells (44 (link)) were purchased from the American Type Culture Collection and cultured in Iscove's modified Dulbecco's medium (Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS), 100 units/ml penicillin, and 100 μg/ml streptomycin (Merck). HUVECs were purchased from Cell Applications and grown in endothelial cell basal medium 2 supplemented with SupplementPack Endothelial Cell GM2 (PromoCell) on collagen-coated dishes. MEDEP-E14 cells (45 (link)) were obtained from RIKEN BioResource Research Center and grown in Iscove's modified Dulbecco's medium containing 15% FBS, 100 units/ml penicillin, 100 μg/ml streptomycin, 1× ITS Liquid Media Supplement (Merck), 0.45 mM α-monothioglycerol (Merck), 50 mg/ml ascorbic acid (Merck), and 3 units/ml human erythropoietin (Kyowa Kirin). Cells were cultured at 37 °C under 5% CO2. Transfections were performed using the reagents Lipofectamine and PLUS (Thermo Fisher Scientific), according to the manufacturer's instructions.
10
Culturing Human Hepatocytes and Endothelial Cells
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Human normal CL-48 hepatocytes were obtained from (Manassas, VA, USA). The cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with nonessential amino acids, l -glutamine, a 2× vitamin solution (Life Technologies Inc., Grand Island, NY, USA), sodium pyruvate, 10% fetal bovine serum, penicillin, and streptomycin (Flow Labs, Rockville, MD, USA). HUVECs were obtained from Cell Applications (San Diego, CA, USA) and then maintained in endothelial cell growth medium (Cell Applications). HUVECs were used at no more than passage 5. All of the cells were cultured at 37 °C in a humidified atmosphere of 5% CO2 and 95% air.
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