Human umbilical vein endothelial cells (HUVECs) and human saphenous vein endothelial cells (HSaVECs) were obtained from Promocell. Cells were cultured in endothelial cell growth medium (Promocell; C-22010) or endothelial cell basal medium (Promocell; C-22210) supplemented with 100 μg/ml penicillin and 100 IU/ml streptomycin; basal medium was additionally supplemented with 0.5–2.0% FCS. Additional components of the complete endothelial cell growth medium included 20 μl/ml foetal calf serum, 4 μl/ml endothelial cell growth supplement, 0.1 ng/ml recombinant human epidermal growth factor, 1 ng/ml recombinant human bFGF, 90 μg/ml heparin and 1 μg/ml hydrocortisone.
C 22210
Manufactured by PromoCell
The C-22210 is a laboratory centrifuge designed for general-purpose applications. It features a compact and robust design, accommodating samples with a maximum volume of 50 mL. The unit operates at a maximum speed of 6,000 rpm, generating a maximum relative centrifugal force of 4,500 x g. Its temperature control range is from 4°C to 40°C.
Automatically generated - may contain errors
Lab products found in correlation
C 22010, by PromoCell (2 mentions)
Huvecs, by PromoCell (2 mentions)
Hsavecs, by PromoCell (1 mentions)
Icrt14, by Bio-Techne (1 mentions)
Hcaec, by PromoCell (1 mentions)
C 12221, by PromoCell (1 mentions)
C 22220, by PromoCell (1 mentions)
C 12203, by PromoCell (1 mentions)
C 22020, by PromoCell (1 mentions)
M3148, by Merck Group (1 mentions)
Calcein am, by AAT Bioquest (1 mentions)
Vascular endothelial growth factor (vegf), by Merck Group (1 mentions)
C 39210, by PromoCell (1 mentions)
Everolimus, by Novartis (1 mentions)
5 protocols using c 22210
1
Isolation and Culture of Human Vascular Endothelial Cells
2
Comparison of HUVEC and HCAEC Responses
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Human umbilical vein endothelial cells (HUVECs, pooled from up to four different donors per lot) and human coronary artery endothelial cells (HCAECs, single donor per lot) from different donors were purchased from Promocell (C-12203 and C-12221), and were utilized between passages 2 and 6. Experiments were repeated with endothelial cells from different lots. HUVECs and HCAECs were cultured in endothelial cell growth medium (Promocell; C-22010 and C-22020) or 2% (v/v) FBS endothelial cell basal medium (Promocell; C-22210 and C-22220) supplemented with 100 mg/mL penicillin and 100 IU/mL streptomycin. Human recombinant TNF-α protein was procured from R&D Systems (210-TA-20). Pharmacological inhibitor of β-catenin, iCRT-14, was acquired from Tocris Bioscience (4299). Cells were maintained in a humified atmosphere with 5% CO2 at 37°C.
3
Culturing Adult Dermal Fibroblasts and Endothelial Progenitor Cells
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Adult HDFs (HDFa), HDFa16, HDFa75 (Thermo Fisher Scientific, C0135C), and BJ foreskin fibroblast (ATCC, CRL-2522) were cultured in DMEM high glucose (Merck, D5546) with FBS (10%, Merck, F0804), L-glutamine (2 mM, Thermo Fisher Scientific, 25030024) and 2-mercaptoethanol (100 μM, Merck, M3148) on 0.1% gelatin-coated plates. Peripheral blood-derived EPCs (C26b, EPC1, and EPC2) were cultured as described (Ormiston et al., 2015 (link)) in endothelial cell basal medium (PromoCell, c-22210) supplemented with 10% FBS and cytokines, without heparin.
4
Adhesion of Monocytes to Endothelial Cells
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HUVEC were cultured at 2 × 104 cells/well in 48-well plates until confluent, then starved with endothelial basal medium (Promocell, C-22210) containing 0.5% fetal bovine serum (FBS) for 6 h, before treatment with TNFα (100 ng/ml), TNFα (100 ng/ml) + NSA (4 μM), TNFα (100 ng/ml) + TPCA-1 (5 μM) or TNFα (100 ng/ml) + Nec-1s (10 μM) for 12 h. Then, 2 × 105 iBMDM pre-labeled with calcein-AM (4 μM, AAT Bioquest, 22003) were seeded on the HUVEC monolayer and allowed to adhere for 30 min. Non-adherent iBMDM were washed away with 200 μl phosphate-buffered saline (PBS) per well three times, and the number of adherent iBMDM was imaged and quantified.
5
Patupilone and Everolimus Sensitivity Study
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Patupilone (epothilone B, EPO906) and Everolimus (RAD001) were provided by Novartis Pharma (Basel, Switzerland). For in vitro experiments, patupilone and Everolimus were dissolved in DMSO (1 mM stock solution) and further diluted in serumcontaining medium. VEGF (Sigma, #V7259) was dissolved in H 2 O (1 mg/mL) and further diluted in serum-containing medium. A549, A549EpoB40, and SW480 cells were grown in RPMI 1640 containing 10% fetal bovine serum, 1% penicillin-streptomycin, and 2 mM L-glutamine at 37°C in 5% CO 2 . Genotyped A549 and A549EpoB40 cells were a gift from Susan Band Horwitz (Albert Einstein College of Medicine, NY) and phenotypically tested for patupilone sensitivity. SW480 cells were obtained from ATCC. Human umbilical vein cells (HUVECs) were cultured in endothelial growth medium (Promo Cell, #C-22210), supplemented with growth factors (PromoCell, C-39210#), 1% penicillin-streptomycin at 37°C in 5% CO 2.
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