Human ECs were purchased from different vendors to capture a broad spectrum of sampling. Specifically, we obtained HUVECs (C2519A; Lonza; C-12203; PromoCell), HSaVECs (HSVEC/A; VEC-Technologies; cAP-0019; Angio-Proteomie), HAECs (PCS-100-011; ATCC; 6100; ScienCell), HIAECs (CC-2545; Lonza; cAP-0020; Angio-Proteomie), HBMVECs (ACBRI 376 V; Cell-Systems; cAP-0002; Angio-Proteomie), HUMVECs (C-12295; PromoCell; 7000; ScienCell), HLMVECs (3000; ScienCell; C-12281; PromoCell), HAMVECs (7200; ScienCell), and HDMECs (C-12212; PromoCell; 2000; ScienCell). All types of ECs were cultured in EGM-2 endothelial growth medium (CC-3162; Lonza) supplemented with 5% FBS (Omega Scientific) and were maintained at 37°C with 5% CO2. Similar EC passages 4–6 were used for final experiments. Human dermal fibroblasts were a gift from W. Lowry (University of California, Los Angeles, Los Angeles, CA), expanded in DMEM supplemented with 10% FBS, and cultured in EMG-2 medium over 48 h before RNA expression experiments. Brightfield images were acquired by using an Axiovert 200M inverted microscope (Zeiss).
Pcs 100 011
Manufactured by American Type Culture Collection
Sourced in United States
The PCS-100-011 is a cell culture incubator designed for optimal growth and maintenance of various cell lines. It provides a controlled environment with temperature, humidity, and CO2 regulation to support cell culture operations.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
C 12281, by PromoCell (1 mentions)
Cc 3162, by Lonza (1 mentions)
Axiovert 200m inverted microscope, by Zeiss (1 mentions)
C 12212, by PromoCell (1 mentions)
Cc 2545, by Lonza (1 mentions)
C 12203, by PromoCell (1 mentions)
Fetal bovine serum (fbs), by Omega Scientific (1 mentions)
Fb 11, by Omega Scientific (1 mentions)
Mcdb131 wofbs, by Omega Scientific (1 mentions)
C57bl 6 mouse primary aortic endothelial cells, by Cell Biologics (1 mentions)
C57 6052, by Cell Biologics (1 mentions)
L glutamine, by Cell Biologics (1 mentions)
Endothelial cell medium supplement kit, by Cell Biologics (1 mentions)
Vascular endothelial growth factor (vegf), by Cell Biologics (1 mentions)
Antibiotic antimycotic solution, by Cell Biologics (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Mitoebselen 2, by MedKoo Biosciences (1 mentions)
4 protocols using pcs 100 011
1
Endothelial Cell Culture and Characterization
2
Culturing Primary Endothelial Cells
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HUVECs (VEC Technologies #HUVEC/T-75 Flask), HAECs (Lonza #CC-2535 and ATCC # PCS-100–011), and BAECs (isolated from bovine aortas) were cultured in complete MCDB-131 medium (VEC Technologies; MCDB131-WOFBS) with 10% fetal bovine serum (Omega Scientific #FB-11). All primary endothelial cells were checked for mycoplasma and cultured on gelatin (Fisher Scientific # NC9369923) coated dishes in humidified incubator at 37 °C and 5% CO2 and used between passages 4 and 9.
3
In Vitro Cytotoxicity Assay on Endothelial Cells
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Two different cell lines, C57BL/6 mouse primary aortic endothelial Cells (Cellbiologics, USA) and human aortic endothelial cells (ATCC®, USA), were used for in vitro cytotoxicity testing. C57BL/6 mouse primary aortic endothelial Cells were purchased at Cellbiologics (C57-6052, Lot#M111718W10, Chicago, Illinois, USA) and human aortic endothelial cells were purchased by ATCC® (PCS-100-011, Lot#64408088, Manassas, Virginia, USA).
The cells grew up in T25 and T75 tissue culture flasks (Falcon™) with endothelial cell medium, which was supplemented with endothelial Cell Medium supplement Kit (0.5 mL VEGF, 0.5 mL ECGS, 0.5 mL heparin, 0.5 mL EGF, 0.5 mL hydrocortisone, 5.0 mL L-glutamine, 5.0 mL antibiotic-antimycotic solution and 25.0 mL FBS; Cellbiologics, USA). When cells were confluent, they were seeded in 6-well tissue culture plates or 96-well tissue culture plates. If cells were 80% confluent, they were used for experiments.
The cells grew up in T25 and T75 tissue culture flasks (Falcon™) with endothelial cell medium, which was supplemented with endothelial Cell Medium supplement Kit (0.5 mL VEGF, 0.5 mL ECGS, 0.5 mL heparin, 0.5 mL EGF, 0.5 mL hydrocortisone, 5.0 mL L-glutamine, 5.0 mL antibiotic-antimycotic solution and 25.0 mL FBS; Cellbiologics, USA). When cells were confluent, they were seeded in 6-well tissue culture plates or 96-well tissue culture plates. If cells were 80% confluent, they were used for experiments.
4
HAEC Knockdown and Overexpression Assay
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Primary HAECs (#PCS-100-011, ATCC) were purchased and maintained in MCDB 131 medium supplemented with 10% fetal bovine serum (FBS), 2 mmol/L glutamax, 10 U/mL penicillin, 100 μg/ml streptomycin (GIBCO/Life Technologies), 30 μg/ml heparin sodium, and bovine brain extract (25 μg/ml). For Ng knockdown and overexpression, HAECs at 70% confluency were transfected with siRNA (20 pmol, #s9723, Thermo Fisher) or plasmid DNA vector targeting Ng overexpression with an HA tag (2 μmol, VectorBuilder ID #VB211018-1249cnr), respectively, using Lipofectamine RNAiMAX (#13778150, Thermo Fisher) for 8 h. Cells were then switched to growth media and experiments performed 24 h later. When applicable, before experiments, transfected cells were treated with 1 nmol MitoEbselen-2 (#564356, MedKoo Biosciences) in reduced-serum media for 30 min.
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