Neuroblastoma cell lines SK-N-BE(2) were purchased from the European Collection of Authenticated Cell Cultures (ECACC; Cat.No. 95011815) and cultured in RPMI-1640 Medium (Sigma, R8758) supplemented with 10% (v/v) FBS and 1% penicillin/streptomycin (RPMI expansion medium). Cells were cultured at 37°C and 5% CO2 in a humidified incubator. Human Umbilical Vein Endothelial Cells (HUVEC) were purchased from Lonza and cultured according to the manufacturer's specifications. HUVEC cells were cultured in HUVEC expansion medium EBM-2 Basal Medium (Lonza CC-3156) supplemented with EGM-2 BulletKit (Lonza CC-3156 & CC-4176) at 37°C in a humidified incubator at 5% CO2.
Human umbilical vein endothelial cells (huvec)
Manufactured by Lonza
Sourced in United States, Switzerland, Belgium, Germany, United Kingdom, Italy, Spain
HUVEC is a primary cell line derived from human umbilical vein endothelial cells. It is commonly used in cell culture research to study endothelial cell biology and function.
Automatically generated - may contain errors
Lab products found in correlation
Egm 2, by Lonza (44 mentions)
Egm 2 bulletkit, by Lonza (33 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (28 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (18 mentions)
Huvecs, by Lonza (17 mentions)
Human umbilical vein endothelial cells (huvec), by Thermo Fisher Scientific (15 mentions)
Ebm 2, by Lonza (14 mentions)
Egm 2mv, by Lonza (13 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (13 mentions)
Egm 2 medium, by Lonza (13 mentions)
L glutamine, by Thermo Fisher Scientific (13 mentions)
Streptomycin, by Thermo Fisher Scientific (10 mentions)
Fetal bovine serum (fbs), by Merck Group (10 mentions)
Penicillin, by Thermo Fisher Scientific (9 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (9 mentions)
Human umbilical vein endothelial cells (huvec), by American Type Culture Collection (9 mentions)
Fetal bovine serum (fbs), by Lonza (8 mentions)
Rpmi 1640, by Thermo Fisher Scientific (8 mentions)
Human umbilical vein endothelial cells (huvec), by Merck Group (8 mentions)
Penicillin streptomycin, by Merck Group (8 mentions)
396 protocols using human umbilical vein endothelial cells (huvec)
1
Culturing Neuroblastoma and HUVEC Cells
2
Characterization and Cultivation of HUVEC
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Human umbilical vein endothelial cells (HUVEC) were purchased from Lonza and cultured in M199 medium (Corning) supplemented with 10% FBS, penicillin–streptomycin, endothelial cell growth factor from sheep brain extract, and 5 U/ml heparin on gelatin–coated dishes in a 37 °C incubator with 5% CO2. Passage 4 – passage 7 cells were used for experiments. Endothelial markers including VE-Cad, CD31, and ERG were used for cell characterization by immunofluorescence.
Detailed cell authentication information of HUVEC can be acquired from Lonza. In brief, routine characterization of HUVEC includes morphological observation throughout serial passages and all lots produced after 01 May 2010 stain at least 90% double-positive for endothelial cell markers CD31/CD105 and negative for alpha smooth muscle actin marker at passage 4. Pre-screened HUVEC are guaranteed to test positive for angiogenesis and vascular cell health markers, eNOS, Tie-2, VEGFr2, and Axl in the first passage out of cryopreservation (From Lonza).
Upon purchase, all cells are performance assayed and test negative for HIV-1, mycoplasma, hepatitis-B, hepatitis-C, bacteria, yeast and fungi. Cell performance and characterization are measured after recovery from cryopreservation.
Detailed cell authentication information of HUVEC can be acquired from Lonza. In brief, routine characterization of HUVEC includes morphological observation throughout serial passages and all lots produced after 01 May 2010 stain at least 90% double-positive for endothelial cell markers CD31/CD105 and negative for alpha smooth muscle actin marker at passage 4. Pre-screened HUVEC are guaranteed to test positive for angiogenesis and vascular cell health markers, eNOS, Tie-2, VEGFr2, and Axl in the first passage out of cryopreservation (From Lonza).
Upon purchase, all cells are performance assayed and test negative for HIV-1, mycoplasma, hepatitis-B, hepatitis-C, bacteria, yeast and fungi. Cell performance and characterization are measured after recovery from cryopreservation.
3
Evaluating hCM and HUVEC Proliferation
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All experiments were performed with the approval of the Institutional Biosafety Committee, National University of Singapore (NUS) under protocol 2018-00717. hCM (Passage 4, Promocell, Heidelberg, Germany) and HUVEC (Passage 4, Lonza, Basel, Switzerland) were cultured according to suppliers’ protocols for approximately 3–4 days in the recommended growth media (hCM—myocyte growth medium, Promocell, Heidelberg, Germany; HUVEC—endothelial growth medium EGM-2, Lonza, Basel Switzerland) and cell seeding density (hCM—10,000 cells/cm2; HUVEC—5,000 cells/cm2). For expansion, the cells were maintained in an incubator at 37 °C with stable 5% CO2 in normoxic conditions with two-daily medium change. hCM and HUVEC were washed with phosphate buffered saline buffer (PBS, Gibco, Carlsbad, CA, USA) or HEPES buffered saline solution (HBSS, Lonza, Basel, Switzerland) respectively, trypsinized and harvested according to the suppliers’ instructions. hCM and HUVEC were cultured in 96-well plates at the recommended cell seeding density (n = 5 for oxygen condition, cell culture media and individual time-points) in preparation for proliferation assays. Cells were maintained at 37 °C with stable 5% CO2 in normoxia or hypoxia. Cells were cultured in 100μL of (1) recommended growth media, (2) basal MEMα, (3) normoxic AFSC secretome (nAFSC-S) or 4) hypoxic AFSC secretome (hAFSC-S).
4
Culturing Common Cell Lines for Research
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HMEC-1: Immortalized human micro vascular endothelial cells (ATCC). HUVEC: Human umbilical vein endothelial cells (Lonza). HMEC-1 and HUVEC cells were cultured in Endothelial Growth Medium (Lonza). hMSC: Immortalized human bone marrow stromal cells44 (link), kindly supplied by Professor Suresh Rattan, Department of Molecular biology and Genetics at Aarhus University. hMSC cells were cultured in DMEM with 10% FCS. HEK 293: Human embryonic kidney cells (ATCC, USA). HEK293 cells were cultured in DMEM with 10% FCS.
ASF-2: Primary human skin fibroblast cells. ASF-2 cells were cultured in DMEM with 10% FCS. ASF-2 cells were kindly supplied by Professor Suresh Rattan Institute of Molecular biology and Genetics at Aarhus University. MCF-7: Human luminal epithelial breast cancer cell line (ATCC). MCF-7 cells were cultured in DMEM with 10% FCS and 0.01 mg/mL insulin.
All plasticware used for cell culturing were from Corning. All cells were grown at 37 °C with 5% CO2 and high humidity and passaged when 80% confluent.
ASF-2: Primary human skin fibroblast cells. ASF-2 cells were cultured in DMEM with 10% FCS. ASF-2 cells were kindly supplied by Professor Suresh Rattan Institute of Molecular biology and Genetics at Aarhus University. MCF-7: Human luminal epithelial breast cancer cell line (ATCC). MCF-7 cells were cultured in DMEM with 10% FCS and 0.01 mg/mL insulin.
All plasticware used for cell culturing were from Corning. All cells were grown at 37 °C with 5% CO2 and high humidity and passaged when 80% confluent.
5
Colorectal Cancer Cell Line Cultivation
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Human colorectal cancer (CRC) cell lines KM12-SM and HCT116 were purchased from the American Type Culture Collection (Manassas, VA, USA). KM12-SM was established from liver metastasis spices that were generated by KM12 injection in nude mouse [29 (link)]. KM12 was established from primary human colorectal carcinoma [29 (link)]. HCT116 was established from human colorectal carcinoma. The normal human dermal fibroblast cell (NHDF), human umbilical vein endothelial cell (HUVEC), and endothelial growth medium (EGM-2MV) were purchased from Lonza (Basel, Switzerland). GFP-expressed HUVEC was purchased from Angio-Proteomie (Boston, MA, USA). RFP-expressed HT29 were purchased from Anticancer (San Diego, CA, USA). HT29 is a human colorectal adenocarcinoma cell line. KM12-SM, HCT116, NHDF and HT29-RFP were cultured in Dulbecco modified Eagles’s medium (DMEM) High-Glucose (Nacalai tesque, 08458, Kyoto, Japan) with antibiotics (Nacalai tesque, 02894, Kyoto, Japan), supplemented with 10% FBS at 37 °C and 5% CO2 in a humidified incubator. HUVEC and HUVEC-GFP were cultured in endothelial growth medium (EGM-2MV, Lonza CC4147, Basel, Switzerland) supplemented with 10% FBS at 37 °C at 5% CO2 in a humidified incubator.
6
Cell Lines for Cancer Research
7
Cell Culture Protocols for Cell Lines
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A549, HeLa, MDA-MB-231 and MCF7 cells were obtained from the American Type Culture Collection (ATCC, VA, USA) and HUVEC from Lonza Inc. (Basel, Switzerland). MCF7, HeLa and hybridoma cells were cultured in DMEM (Hyclone, UT, USA) with 10% fetal bovine serum (FBS). A549 and MDA-MB-231 cells were maintained in RPMI (Hyclone, UT, USA) 1640 media containing 10% FBS and HUVEC in EGM-2 (Lonza, Basel, Switzerland).
8
HUVEC Transfection with SMAD6 siRNA
9
Cultivation of Microvascular and Umbilical Vein Endothelial Cells
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Human microvascular endothelial cells (MEC) and human umbilical vein endothelial cells (HUVEC) were purchased from Lonza (Walkersville, MD). MEC were propagated in microvascular endothelial cell growth medium-2 (EGM-2MV, Lonza). HUVEC were grown in endothelial cell growth medium-2 (EGM-2, Lonza). Cells were maintained under standard conditions of 37°C and 5% CO2 and the passage numbers of the endothelial cells were kept between 3 and 6.
10
Cell Culture of Cancer and Endothelial Cell Lines
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HT-29 colon cancer cell line and 2H-11 mouse endothelial cell lines were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA), while HUVEC was purchased from Lonza (Basel, Switzerland). Cells were maintained in a complete medium, either Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies, Carlsbad, CA, USA) containing 10% heat-inactivated fetal bovine serum (FBS) (Corning, NY, USA), Penicillin (100 U/mL), Streptomycin (100 µg/mL) (Life Technologies) and 2 mM L-alanyl-L-glutamine dipeptide (GlutaMAX Supplement; Life Technologies) for HT-29 and 2H-11 or EBM™-2 Endothelial Cell Growth Basel Medium, supplemented with EBM™-2 Endothelial SingleQuots™ kit (Lonza) for HUVEC. Cells were grown under standard culture conditions (37 °C with 5% CO2) and passaged no more than 5 times for HUVEC and no more than 20 times for HT-29 and 2H-11. All cell lines were mycoplasma-free, as determined using the MycoAlert™ Mycoplasma Detection Kit (Lonza) and/or a custom PCR-based assay, as described previously [11 (link),51 (link)].
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