Human umbilical vein endothelial cell (HUVEC) and two murine endothelial cell lines (2H-11 and 3B-11) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). 2H-11 and 3B-11 lines were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Life Technologies, Carlsbad, CA, USA), supplemented with 10% foetal bovine serum (FBS; Corning, Corning, NY, USA), 2 mM L-alanyl-L-glutamine dipeptide (GlutaMAXTM; Life Technologies) and 50 U/mL-50 µg/mL penicillin–streptomycin (Life Technologies). HUVEC line was maintained in Vascular Cell Basal Medium supplemented with Endothelial Cell Growth Kit-VEGF (ATCC). Cultures were maintained in 5% CO2 at 37 °C.
Human umbilical vein endothelial cells (huvec)
Manufactured by American Type Culture Collection
Sourced in United States, China, United Kingdom, Japan, Italy
The HUVEC (Human Umbilical Vein Endothelial Cells) is a primary cell line derived from the human umbilical vein. It is a widely used in vitro model for studying endothelial cell biology and angiogenesis.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (74 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (40 mentions)
Mda mb 231, by American Type Culture Collection (22 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (22 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (19 mentions)
Human umbilical vein endothelial cells (huvec), by Thermo Fisher Scientific (17 mentions)
Mcf 7, by American Type Culture Collection (16 mentions)
Rpmi 1640, by Thermo Fisher Scientific (16 mentions)
Hek293, by American Type Culture Collection (15 mentions)
Penicillin, by Thermo Fisher Scientific (15 mentions)
Hct116, by American Type Culture Collection (13 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (13 mentions)
Streptomycin, by Thermo Fisher Scientific (12 mentions)
Hek293t, by American Type Culture Collection (11 mentions)
Endothelial cell growth kit vegf, by American Type Culture Collection (10 mentions)
Thp 1, by American Type Culture Collection (10 mentions)
Heparin, by Merck Group (9 mentions)
Human umbilical vein endothelial cells (huvec), by Lonza (9 mentions)
Hepg2, by American Type Culture Collection (9 mentions)
Sh sy5y, by American Type Culture Collection (8 mentions)
388 protocols using human umbilical vein endothelial cells (huvec)
1
Endothelial Cell Culture Protocol
2
Cell Line Cultivation and Maintenance
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Molt-4, Jurkat (clone E6-1), LS174T, HL-60, WEHI-3, HUVEC, and human dermal fibroblasts were purchased from ATCC. Lox were a kind gift from Dr. Oystein Fodstad (Oslo University Hospital). MKN45, Colo205, MDA-MB-231, and MCF7 were a kind gift from Dr. Henrik Clausen (University of Copenhagen). Tn(−) and Tn(+) LS174T were subcloned from a mixed population as previously described19 (link). Lox and Jurkat were transfected with full length Cosmc or empty vector (pcDNA3.1+) and selected by G418. Tn(−) cells were further sorted by FACS. Molt-4, Jurkat, Lox, HL-60, MKN45, and Colo205 were cultured in RPMI (Corning) supplemented with 10% FBS and 2% P/S. LS174T, MDA-MB-231, and MCF7 were cultured in DMEM (Corning) supplemented with 10% FBS and 2% P/S. MCF7 were further supplemented with 0.01 mg/ml insulin. WEHI-3 were cultured in Iscove’s (Corning) supplemented with 10% FBS, 0.05 mM 2-ME, and 2% P/S. HUVEC and human dermal fibroblasts were cultured with endothelial cell growth kit-VEGF (ATCC) and fibroblast growth kit- low serum (ATCC) as instructed. All cells were cultured on plastic, except HUVECs, which were cultured on plastic pre-coated with 0.1% gelatin. We did not perform independent verification of cell lines or testing for mycoplasma.
3
Cell Line Authentication and Culture
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MCF-7 human breast cancer cells were obtained from the Michigan Cancer Foundation; BT-474, MDA-MB-231, BT-20 and Human Umbilical Cord Vascular Endothelial Cells (HUVEC) were from the ATCC (Manassas VA). Transformed mouse mammary stromal cells (BJ3Z) were developed in our laboratory [27 (link), 29 (link)]; normal mouse mammary fibroblasts (NMF) were a kind gift of L. Wakefield (NCI) [27 (link), 29 (link)]. All cell lines were authenticated by Single Tandem Repeat analysis at the CU Cancer Center Sequencing Core and were mycoplasma-free. Cells were routinely passaged in minimum essential medium (MEM; Invitrogen, Carlsbad CA) containing 5% fetal calf serum (FCS; HyClone, Logan UT). For estrogen-free conditions the medium was phenol red-free and the serum was stripped of endogenous hormones by two incubations with dextran-coated charcoal (DCC). HUVEC cells were grown in F-12 K medium (ATCC) supplemented with 0.1 mg/ml heparin, 0.05 mg/ml endothelial cell growth supplement (ECGS; Cat N. 356006 BD Biosciences, Bedford, MA) and 10% FCS.
4
Cell Line Cultivation and Maintenance
5
Culture of Chang Liver and HUVEC Cells
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Chang liver cells (human normal cell line) (ATCC, VA, USA) and HUVEC (human umbilical vein endothelial cells) (ATCC, VA, USA) were grown in culture using 25 cm2 polystyrene flasks (Falcon). The Chang liver cells were maintained at 37°C in a humidified 5% CO2 atmosphere and cultured in RPMI medium supplemented with 10% fetal bovine serum, and 1% antibiotic, while the HUVEC were cultured in F-12K medium (ATCC, VA, USA) containing 10% fetal bovine serum (FBS), 1% antibiotic-antimycotic solution, 0.1 mg/ml heparin, and 0.03 mg/ml endothelial cell growth supplemented under an atmosphere of 5% CO2 at 37°C. The cells were subcultured and split 1∶4 every 4 days.
6
Culturing Common Cell Lines for Research
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HEK293, HUVEC, THP-1, and wild-type primary dermal fibroblast cells were purchased from American Type Tissue Collection (ATCC; Manassas, VA, USA). Primary Dermal Fibroblasts cells were grown in Media 106 with the addition of LSGS kit (S-003-10) (ThermoFisher, Rockford, IL, USA) and used between passage 4-10. HEK293 cells were maintained in 5% FBS with Improved Minimum Essential Medium (IMEM) (ThermoFisher, Rockford, IL, USA), THP-1 cells were grown in RPMI (ThermoFisher, Rockford, IL, USA) with 10% FBS following the manufacturer’s recommendation. HUVEC cells were grown in vascular cell basal medium with the addition of VEGF endothelial cell growth kit (ATCC; Manassas, VA, USA), and used between passages 3 and 8.
7
Angiogenic Potential of Transfected Cancer Cells
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Following transfection, the cancer cells were co-cultured with human umbilical vein endothelial cell (HUVEC) (ATCC, Manassas, VA, U.S.A.). Each group was set with three duplicated wells. A total of 60 μl Matrigel was added into each well of a 96-well plate which was gently shaken to enable the Matrigel spread evenly. After the Matrigel gel was coagulated, the HUVEC (ATCC, Manassas, VA, U.S.A.) were resuspended with the supernatant of the transfected HONE-1 cells, with cell concentration adjusted to 2 × 105 ml−1. Afterwards, the above-mentioned cell suspension (150 μl) was cultured in the 96-well plate coated with Matrigel at 37°C with 5% CO2 for 24 h. Then, the tube formation of HUVEC was observed under a phase contrast microscope (×100). Five visual fields were randomly selected from each group to assess the angiogenesis of HUVEC co-cultured with cancer cells in each group, which was expressed as the number of tube formation.
8
Culturing Primary Human Aortic Endothelial Cells
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Primary human aortic endothelial cells (HAEC, ScienCell) were plated on culture dishes pre-coated with 10 ng/ml fibronectin (Millipore) and cultured in endothelial cell medium (ECM, ScienCell) supplemented with 5% fetal calf serum (FCS), 1% endothelial cell growth supplement (ECGS), 100 units/ml penicillin, and 100 µg/ml streptomycin [40] (link). Cells were used from passages 3 to 6 in all experiments. Immortalized human umbilical vein endothelial cells (HUVEC), monocyte cell line THP1, and cervical cancer cell line Caski were purchased from ATCC (Manassas, VA) and cultured in DMEM (HUVEC, Caski), or RPMI 1640 (THP1) containing 10% FCS and antibiotics. All cells were cultured in a humidified atmosphere with 5% CO2 at 37°C.
9
Cell Culture of Liver Cancer and Endothelial Cells
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Human HCC cell lines SNU-398 and SK-HEP-1, and Human umbilical vein endothelial cell (HUVEC) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Human HCC cell line Huh7 was purchased from National Collection of Authenticated Cell Cultures of Chinese Academy of Sciences (Shanghai, China). SNU-398 was maintained in RPMI 1640 medium (Invitrogen, Carlsbad, CA USA) added with 10% fetal bovine serum (FBS, Invitrogen). SK-HEP-1 was maintained in Eagle’s Minimum Essential Medium (Invitrogen) added with 10% FBS. HUVEC was maintained in Vascular Cell Basal Media (ATCC) supplemented with Endothelial Cell Growth Kit (ATCC). Huh7 was maintained in Dulbecco’s modified Eagle’s medium (Invitrogen) added with 10% FBS. All cells were cultured at 37 °C containing 5% CO2 under normoxia. Under hypoxia, cells were cultured at 37 °C containing 1% O2, 94% N2, and 5% CO2. The cells were authenticated using STR profiles. All cells were routinely tested as mycoplasma-free.
10
Cultivation of Human Umbilical Vein Endothelial Cells
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Human primary umbilical vein endothelial cell (HUVEC) and human umbilical vein/vascular endothelial cell line (HUVEC-C) were purchased from ATCC (USA). HUVEC cells were cultured in vascular cell basal medium (ATCC, USA) and supplemented with endothelial Cell Growth Kit (ATCC, USA), and the HUVEC-C cells cultured in basal medium (ATCC, USA) containing 10% FBS (Gibco, USA).
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