Primary cultures of human umbilical vein endothelial cells (HUVECs) were prepared as described previously.18 (link) HUVECs purchased from Bioresource Collection and Research Center (BCRC) in Taiwan were maintained in Medium 199 (Invitrogen, MA, USA) complemented with 10% fetal calf serum, 30 µg/mL endothelial cell growth supplement (R&D Systems, MN, USA), 1% penicillin–streptomycin, and 7.5 U/mL heparin under standard cell culture conditions (humidified atmosphere, 5% CO2, 37°C). Cells between passages 2 and 5 were used for the experiment. In addition, cultures of HL-1 cardiomyocytes were prepared as described previously.19 (link) HL-1 cardiomyocytes (a kind gift from Dr. C. T. Tsai, Division of Cardiology, Department of Internal Medicine and Cardiovascular center, National Taiwan University College of Medicine and Hospital, Taipei City, Taiwan) were cultured on Claycomb medium supplemented with 1 μM retinoic acid, 10 μM norepinephrine (Sigma-Aldrich), 100 units/mL penicillin, 100 μg/mL streptomycin and an additional 1× nonessential amino acids (Life Technologies, CA, USA). The medium was changed approximately every 24 hr. The cells were grown at 37°C in an atmosphere of 5% CO2 and 95% air at a relative humidity of approximately 95%.
Huvecs
Manufactured by RIKEN BioResource Center
Sourced in United States
Human umbilical vein endothelial cells (HUVECs) are primary endothelial cells derived from the umbilical vein. They are commonly used in in vitro studies of angiogenesis, vascular biology, and related fields.
Automatically generated - may contain errors
Lab products found in correlation
Huvecs, by Merck Group (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Endothelial cell growth supplement (ecgs), by Merck Group (3 mentions)
Endothelial cell medium (ecm), by ScienCell (3 mentions)
Bhk 21, by Japanese Collection of Research Bioresources Cell Bank (2 mentions)
C6 36, by Japanese Collection of Research Bioresources Cell Bank (2 mentions)
Egm 2, by Lonza (2 mentions)
Kanamycin, by Merck Group (2 mentions)
Heparin, by Merck Group (2 mentions)
Endothelial cell medium (ecm), by Lonza (2 mentions)
Medium 199, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Collagen type 1, by Corning (1 mentions)
Egm 2 endothelial growth medium, by Lonza (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Endothelial cell growth supplement (ecgs), by R&D Systems (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
32 protocols using huvecs
1
Isolation and Culture of Human Endothelial and Cardiac Cells
2
HUVEC and Cardiac Fibroblast TGF-β Modulation
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Primary human umbilical vein endothelial cells (HUVECs) were purchased from Bioresource Collection and Research Center, Hsinchu, Taiwan. The HUVECs were cultured in M199 medium containing 5% fetal bovine serum (FBS), 1% endothelial cell growth factors (Millipore 02-102) and 1% penicillin/streptomycin in 5% CO2 at 37 °C. Only HUVECs at 3–6 passages were used for this experiment. The cells were cultured in 0.1% gelatin coating monolayer dishes. At approximately 80% confluence, the culture medium was changed to a serum-free solution for 24 h before the use of the cells in the experiments. The primary mouse cardiac fibroblasts were isolated from a pathogen-free neonate C57BL/6 mouse and cultured on pre-coated gelatin-based culture dishes as described in the previous study [31 (link)]. To examine the effect of TGF-β, the HUVECs or cardiac fibroblasts were treated with PBS, TGF-β1 (10 ng/mL) +10 ng TGF-β2 (10 ng/mL) or TGF-β1 (10 ng/mL) +10 ng TGF-β2 (10 ng/mL) with calcitriol (100 ng/mL) for 72 h according to the previous study [32 (link)]. Cell morphology, protein expression and gene expression were analyzed the EndMT markers using Western blot and immunofluorescent staining.
3
Culturing Human Cell Lines
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Human CCA cell lines were purchased from Korean Cell Line Bank (Seoul, Korea). Cells were grown in RPMI 1640 medium supplemented with 10% FBS and 1% antibiotic-antimycotic agents. Culture medium was changed thrice per week. Human vascular endothelial cells (HUVECs) were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan R.O.C.) and maintained as previously described.14 (link)
4
Human Umbilical Vein Endothelial Cell Culture
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Primary human umbilical vein endothelial cells (HUVECs) were purchased from the Bioresource Collection and Research Center (BCRC), Hsinchu, Taiwan. Briefly, cells were cultured in a M199 medium containing 5% fetal bovine serum (FBS), 1% endothelial cell growth factors (02-102, Millipore, Burlington, MA, USA), and 1% penicillin/streptomycin at 37 °C with 5% CO2. HUVECs between 3 to 6 passages were cultured in monolayer dishes for the experiment. At approximately 80% confluence, the culture medium was changed to a serum-free medium for 24 h before the cells were used for further experiments.
5
HUVEC Culture Using Collagen Coating
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HUVECs (Bioresource Collection and Research Center, Hsinchu, Taiwan) were used between passages 4 and 9 and maintained in EGM®-2 endothelial growth medium (CC-3162, Lonza, Basel, Switzerland) in T75 tissue culture flasks precoated with 50 µg ml−1 collagen coating solution for 30 min at 37°C to foster cell adhesion and growth (Park et al., 2013 (link)). The collagen coating solution was prepared using collagen type I (3.61 mg ml−1, Corning, NY, United States) and diluted in 0.02 N acetic acid (Sigma-Aldrich, MO, United States).
6
Cultivating and Characterizing Five HCC Cell Lines
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Five HCC cell lines, Huh7, Hep3B, PLC/PRF/5, SK-Hep-1, and HepG2, were used. Huh7 was obtained from the Japanese Collection of Research Bioresources. Hep3B, PLC/PRF/5, and SK-Hep-1 were purchased from the American Type Culture Collection. HepG2 cells and HUVECs were obtained from the Bioresource Collection and Research Center, Taiwan. Hep3B cells were maintained in Eagle’s minimum essential medium supplemented with 10 % heat-inactivated fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin. The other HCC cells were grown in Dulbecco’s minimum essential medium supplemented with 10 % FBS, 100 mM nonessential amino acids, 100 U/mL penicillin, and 100 μg/mL streptomycin. All cells were cultured in a humidified incubator at 37 °C and 5 % CO2. All reagents for cell culture were purchased from Life Technologies (Grand Island, NY, USA).
7
Hyperglycemia Effects on HUVEC Cells
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Human umbilical vein endothelial cells (HUVECs) were obtained from the Bioresource Collection and Research Center (BCRC), Hsinchu, Taiwan. HUVECs were grown in M199 medium supplemented with 10% FBS, 30 µg/mL ECGS, 25 U/mL heparin, 2 mM L-glutamine, 1.5 g/L sodium bicarbonate and 100 U/L penicillin and streptomycin at 37°C in a humidified atmosphere of 5% CO2. Hyperglycemia treatment was induced by treating HUVECs with 15, 30 and 60 mM of d-glucose for 24-72 h. HUVEs were also incubated with HG in the presence or absence of ADC (10 µM) or resveratrol (5 µM). Controls were performed in the presence of media with normal glucose (NG, 5.5 mM).
8
Cell Culture Protocols for Various Cell Lines
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HUVECs (Bioresource Collection and Research Center, Taiwan) were cultured in EGM-2 (Lonza, Basel, Switzerland), and the human monocytic cell line THP-1 (Bioresource Collection and Research Center, Taiwan) was cultured in Roswell Park Memorial Institute 1640 Medium (RPMI 1640; Thermo Fisher Scientific, Waltham, MA, USA). The baby hamster kidney cell line BHK-21 and Aedes albopictus cell line C6/36, purchased from the Japanese Collection of Research Bioresources (JCRB Cell Bank, Japan) and the American Type Culture Collection (ATCC, Manassas, Virginia, USA), were maintained in Dulbecco’s modified Eagle’s medium (DMEM). The medium used to grow all cell types was supplemented with 10% fetal bovine serum (FBS; HyClone Laboratory, Logan, UT, USA). All cells were cultured at 37°C in a 5% CO2 atmosphere except for C6/36 cells, which were cultured at 28°C.
9
High-Glucose Treatment of Fibroblasts and Endothelial Cells
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Human normal dermal fibroblasts (HNDFs, CCD966SK) and human umbilical vein endothelial cells (HUVECs) were obtained from the Bioresource Collection and Research Center (BCRC), Hsinchu, Taiwan. HNDFs were grown in MEM containing 10% FBS, 2 mM L-glutamine, 100 U/mL penicillin and streptomycin at 37°C in a fully humidified atmosphere of 5% CO2. Likewise, HUVECs were grown in M-199 medium supplemented with ECGS, heparin, 10% FBS and 100 U/mL penicillin and streptomycin at 37°C in a fully humidified atmosphere of 5% CO2. High-glucose treatment was performed by treating cells with 15 or 30 mM D-glucose (HG) for 24-72 h. HNDFs and HUVECs were also treated with HG in the presence of 10 μM ANM or 100 μM N-acetylcysteine or 5 μM resveratrol. Controls were performed in the presence of media with normal glucose alone (NG, 5.5 mM) or with 10 μM ANM or 100 μM N-acetylcysteine or 5 μM resveratrol.
10
Establishing Sarcoma and Endothelial Cell Lines
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Human uterine sarcoma cell line MES-SA was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). MES-SA/Dx5 is a multidrug-resistant subline derived from MES-SA. Human umbilical vein endothelial cells (HUVECs) were procured from the Bioresource Collection and Research Center, Food Industry Research and Development Institute (BCRC, Hsinchu, Taiwan). MES-SA/Dx5 cells were maintained in McCoy’s 5A medium supplemented with 10% FBS, 1 mM sodium pyruvate, 100 U/mL penicillin, 100 μg/mL streptomycin and 0.4 μg/mL of DOX. HUVECs were grown in M199 medium containing endothelial growth medium (EGM)-2 SingleQuots Kit (consisting of FBS, VEGF and other growth factors) (Lonza, Basel, Switzerland), 1 mM sodium pyruvate, 10 mM HEPES, 100 U/mL penicillin and 100 μg/mL streptomycin. All the cells were cultured at 37 °C in a humidified incubator with 95% air and 5% CO2.
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