HASCs were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). HUVECs were purchased from the China Infrastructure of Cell Line Resources (Beijing, China). Isolation of HAMSCs was performed following the pancreatin/collagenase digestion method[23 (link)–25 (link)]. HASCs and HAMSCs were cultured in 60-mm plates in αMEM supplemented with 100 U/L penicillin, 100 mg/L streptomycin, and 10% FBS in a humidified atmosphere of 5% CO2 at 37°C. HUVECs were cultured in 60-mm plates in EBM containing 1% FBS and EGM-2 BulletKit in a humidified atmosphere of 5% CO2 at 37°C. Cells from passage 3 were used and culture medium was changed every 3 days. The study protocols were approved by the Ethics Committee of the School of Stomatology, Nanjing Medical University, China (NO.PJ2013-037-001). Informed consent was obtained from all the participants enrolled in this study.
Huvecs
Manufactured by China Infrastructure of Cell Line Resources
Sourced in China
HUVECs (Human Umbilical Vein Endothelial Cells) are primary cell lines derived from the human umbilical vein. They are a widely used in vitro model for studying endothelial cell biology and function.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
2 hydroxy 4 2 hydroxyethoxy 2 methylpropiophenone irgacure 2959, by Merck Group (1 mentions)
Glutamax 1, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Nih3t3 cells, by American Type Culture Collection (1 mentions)
Catalase, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dispase, by Thermo Fisher Scientific (1 mentions)
10 cm culture dishes, by Corning (1 mentions)
M2 medium, by PromoCell (1 mentions)
Hepg2 cells, by China Infrastructure of Cell Line Resources (1 mentions)
Endothelial cell growth supplement ecgs, by ScienCell (1 mentions)
Penicillin, by ScienCell (1 mentions)
Streptomycin, by ScienCell (1 mentions)
Hbm mscs, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Endothelial cell medium (ecm), by ScienCell (1 mentions)
Hbm mscs, by American Type Culture Collection (1 mentions)
6 protocols using huvecs
1
Isolation and Culture of Human Stem Cells
2
Anti-HPV Protein Hydrogel Fabrication
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Anti-HPV protein (bovine β-lactoglobulin modified with 3-hydroxyphthalic anhydride) was supplied by Jingbo Biological Pharmaceutical Co., Ltd. (Taiyuan, China). A stock solution was prepared in deionized (DI) water at a concentration of 26.9 mg/mL. 2-Hydroxy-4-(2-hydroxyethoxy)-2-methylpropiophenone (Irgacure 2959) and [2-(acryloyloxy)ethyl]trimethylammonium chloride solution (AETAC) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sodium alginate (ALG, (C6H9O7Na)n, Mn = 200,000, G blocks: M blocks = 39:61, low viscosity) was obtained from JiSiEnBei International Trade Co., Ltd. (Hongkong, China). Polyethylene glycol diacrylate (PGEDA, Mn = 6 kDa) was purchased from Huateng Pharmaceutical Co., Ltd. (Changsha, China). NIH 3T3 cells (American Type Culture Collection, Manassas, VA, USA) and human umbilical vein endothelial cells (HUVECs, China Infrastructure of Cell Line Resources, Beijing, China) were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, Grand Island, NY, USA), 1% nonessential amino acid solution (NEAA, Gibco, Grand Island, NY, USA), 1% GlutaMAXTM-I (Gibco, Grand Island, NY, USA), and 1% antibiotics (penicillin and streptomycin, Gibco, Grand Island, NY, USA) at 37 °C in a humidified atmosphere containing 5% CO2.
3
Culturing and Treating HUVECs
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HUVECs were acquired from China Infrastructure of Cell Line Resources (Beijing, China). Cells were grown in Dulbecco's modified Eagle's medium (DMEM, HyClone, USA) containing 10% fetal bovine serum (FBS, HyClone, USA) and 1% penicillin-streptomycin (Gibco, USA). The cell cultures were incubated at 37°C in a humidified atmosphere with 5% CO2. Cells were used for experiments at 80–90% confluency after 2-3 days of culture. In some cases, 5 U/ml catalase (Sigma, USA) was added 5 min before H2O2 was administered.
4
Isolation and Culture of HUVECs and Melanocytes
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HUVECs are purchased from the China Infrastructure of Cell Line Resources (Beijing, China) and were cultured in endothelial basal medium containing 1% fetal bovine serum (FBS) and endothelial cell growth medium 2 Bullet Kit in a humidified atmosphere of 5% CO2 at 37°C. Passages 3 and 5 were used in all experiments.
For melanocytes, adolescent foreskin was obtained from urinary surgery and handled aseptically. After removing subcutaneous elements, the specimen was cut into 0.5 cm2 pieces and put into 0.5% dispase (Gibco, Invitrogen, Auckland, USA) for overnight incubation at 4°C. The epidermis was peeled off from the dermis. The epidermis was further incubated in 0.25% trypsin for 10 min at 37°C. Trypsin activity was neutralized by adding FBS. The mixed cell suspension was filtered through nylon gauze (100 μm mesh) and cells were washed twice with 0.1 M phosphate-buffered saline (PBS) prior to resuspension in M2 medium (PromoCell, Heidelberg, Germany) supplemented with penicillin (100 units/ml) and streptomycin (100 mg/ml). Cells were seeded onto 10 cm culture dishes (Corning, Corning NY, USA), maintained at 37°C in a humidified atmosphere containing 5% CO2. Passages 3 to 5 were used in all experiments, including cell identification by Dopa staining and S-100 protein immunohistochemical staining.
For melanocytes, adolescent foreskin was obtained from urinary surgery and handled aseptically. After removing subcutaneous elements, the specimen was cut into 0.5 cm2 pieces and put into 0.5% dispase (Gibco, Invitrogen, Auckland, USA) for overnight incubation at 4°C. The epidermis was peeled off from the dermis. The epidermis was further incubated in 0.25% trypsin for 10 min at 37°C. Trypsin activity was neutralized by adding FBS. The mixed cell suspension was filtered through nylon gauze (100 μm mesh) and cells were washed twice with 0.1 M phosphate-buffered saline (PBS) prior to resuspension in M2 medium (PromoCell, Heidelberg, Germany) supplemented with penicillin (100 units/ml) and streptomycin (100 mg/ml). Cells were seeded onto 10 cm culture dishes (Corning, Corning NY, USA), maintained at 37°C in a humidified atmosphere containing 5% CO2. Passages 3 to 5 were used in all experiments, including cell identification by Dopa staining and S-100 protein immunohistochemical staining.
5
Cell Culture Conditions for HUVECs and HepG2
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Both HUVECs and HepG2 cells were purchased from the China Infrastructure of Cell Line Resources (Beijing, China) and American Type Culture Collection (Manassas, Virginia), respectively. The HUVECs were cultured in 60-mm plates in endothelial basal medium containing 1% fetal bovine serum (FBS) and endothelial cell growth medium 2 Bullet Kit in a humidified atmosphere of 5% CO2 at 37°C. And HepG2 cells were cultured in Roswell Park Memorial Institute 1640 (RPMI 1640) medium supplemented with 10% FBS, 1% penicillin/streptomycin, and 2% l -glutamine at 37°C in a humidified atmosphere of 5% CO2.
6
Culturing Human Bone Marrow and Umbilical Vein Cells
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Human Bone Mesenchymal Stem Cells (hBM-MSCs) were obtained from the American Type Culture Collection (PTA-1058; ATCC, VA, USA). The hBM-MSCs were cultured in medium including 85% (v/v) α-MEM, 15% (v/v) fetal bovine serum (FBS) (Gibco, Grand Island, NY, USA) and supplemented with 100 U/ml penicillin and 100 µg/ml streptomycin (Gibco, USA). Human Umbilical Vein Endothelial Cells (HUVECs) were obtained from the China Infrastructure of Cell Line Resources (Beijing, China) and cultured in endothelial cell medium (ECM) (#1001, ScienCell Research Laboratories, Inc., CA, USA) containing 10% FBS, Endothelial Cell Growth Supplement (ECGS) (#1052, ScienCell Research Laboratories, Inc., USA), 100 U ml -1 penicillin, and 100 μg ml -1 streptomycin as previously described [20] . Cells from the 3 rd passages to the 6 th passages were also used in the experiment, and the culture medium was changed every three days.
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