Endothelial cell growth kit vegf

Manufactured by American Type Culture Collection

Sourced in United States

The Endothelial Cell Growth Kit-VEGF is a collection of reagents designed to support the growth and proliferation of endothelial cells in cell culture. The kit includes essential components, such as growth factors and supplements, that are specifically formulated to promote the in vitro expansion of endothelial cells.

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Lab products found in correlation

Human umbilical vein endothelial cells (huvec), by American Type Culture Collection (10 mentions) Huvecs, by American Type Culture Collection (8 mentions) Vascular cell basal medium, by American Type Culture Collection (6 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) Rpmi 1640 medium, by Merck Group (4 mentions) Hl 60, by American Type Culture Collection (3 mentions) Trypsin edta, by Thermo Fisher Scientific (3 mentions) Hek293, by American Type Culture Collection (2 mentions) Us qualified fetal bovine serum, by Thermo Fisher Scientific (2 mentions) Wehi 3, by American Type Culture Collection (2 mentions) Fibroblast growth kit low serum, by American Type Culture Collection (2 mentions) Molt 4, by Corning (2 mentions) Jurkat, by Corning (2 mentions) Hl 60, by Corning (2 mentions) Ls174t, by Corning (2 mentions) Iscove s, by Corning (2 mentions) Dulbecco modified eagle medium (dmem), by Corning (2 mentions) Roswell park memorial institute (rpmi), by Corning (2 mentions) Mda mb 231, by Corning (2 mentions)

48 protocols using endothelial cell growth kit vegf

Ang II-induced STAT3 Inhibition

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HCAECs and corresponding growth medium with VEGF endothelial cell growth kit were purchased from ATCC (Manassas, VA). Ang II was purchased from Sigma–Aldrich (Stoughton, MA) and dissolved in phosphate-buffered saline to make 1 mM stock solution. Inhibitor of STAT3 Stattic was purchased from Santa Cruz (Santa Cruz, CA). Anto-mmu-miR21 miScript inhibitor and inhibitor negative control were purchased from Qiagen (Valencia, CA).

Mehta J.L., Mercanti F., Stone A., Wang X., Ding Z., Romeo F, & Khaidakov M. (2015). Gene and MicroRNA Transcriptional Signatures of Angiotensin II in Endothelial Cells. Journal of Cardiovascular Pharmacology, 65(2), 123-129.

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HUVEC Culture and Dexamethasone Treatment

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Normal primary human umbilical vein endothelial cells (HUVECs; Pooled, PCS-100–013) were obtained from ATCC (Manassas, VA, USA) and grown at 37°C and 5% CO₂ on uncoated T75 flasks in vascular cell basal medium (ATCC, PCS-100–030) containing 10% fetal bovine serum (FBS), VEGF endothelial cell growth kit (ATCC, PCS-100–041) and treated with DEX using 2% FBS (both medium were prepared without hydrocortisone hemisuccinate), 100 units/mL penicillin and 100 μg/mL streptomycin. Cells were kept frozen in medium containing 10% DMSO (Bio-Rad Laboratories Canada Ltd., Mississauga, ON, Canada). For experiments, passage 3 was used. Cells were counted using a haemocytometer and seeded onto permeable polyethylene terephthalate (PET) filters at the base of BD Falcon cell culture inserts (BD Biosciences, Mississauga, ON, Canada) at a density of 0.5 × 10⁶ cells/insert.
Dexamethasone (DEX) was obtained from Sigma-Aldrich (Oakville, ON, Canada), and full-length Ad was produced in-house using a mammalian expression system (i.e. HEK 293 cells) according to methods described by (Wang, et al. 2002 (link); Xu, et al. 2004 (link)). HUVECs were treated for 5 days with DEX (1 μM) starting 24 h after seeding cells into inserts, HUVECs were treated with DEX added to both apical and basolateral sides of inserts at concentrations indicated above

Dang T.Q., Yoon N., Chasiotis H., Dunford E.C., Feng Q., He P., Riddell M.C., Kelly S.P, & Sweeney G. (2017). Transendothelial movement of adiponectin is restricted by glucocorticoids. The Journal of endocrinology, 234(2), 101-114.

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HL-60 Derived Exosomes and Neutrophil Elastase Modulate Endothelial Cells

Cited 3 times

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A human HL-60 cell line (ATCC) was cultured in RPMI containing 10% FBS, 1% Primocin, and Insulin-Transferrin-Selenium (ITS-G, 1X, ThermoFisher, Waltham, MA). For differentiation into neutrophil-like cells, the media was supplemented with 1.3% DMSO and the cells grown for 5 days. Cells were then treated with either 1µM fMLP (N-formyl-L-methionyl-L-leucyl-phenylalanine, Sigma) or DMSO overnight in media containing 10% exosome-free FBS. Exosomes were collected from the media by ultracentrifugation.
Human lung microvascular endothelial cells (MVEC) from healthy donors [26 (link)], a gift from Dr. Andrew J. Bryant, were cultured on collagen in Vascular Cell Basal Medium supplemented with the VEGF Endothelial Cell Growth Kit (ATCC, Manassas, VA). Cells were treated with either 1×10⁷ per well of quiescent exosome from HL-60 cells, activated (fMLP treated) exosomes from HL-60 cells [17 (link)], 34 nM of purified neutrophil elastase [27 (link), 28 (link)] (Athens Research and Technology. Athens, GA), 34 nM of neutrophil elastase and purified alpha-1 antitrypsin (Grifols USA, Los Angeles, CA), or activated HL-60 exosomes and alpha-1 antitrypsin (34 nM) overnight before RNA was collected using Trizol.

Lascano J., Oshins R., Eagan C., Wadood Z., Qiang X., Flagg T., Scindia Y., Mehrad B., Brantly M, & Khodayari N. (2022). Correlation of alpha-1 antitrypsin levels and exosome associated neutrophil elastase endothelial injury in subjects with SARS-CoV2 infection. PLoS ONE, 17(9), e0274427.

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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.

Ivanova M.M., Dao J., Kasaci N., Adewale B., Fikry J, & Goker-Alpan O. (2020). Rapid Clathrin-Mediated Uptake of Recombinant α-Gal-A to Lysosome Activates Autophagy. Biomolecules, 10(6), 837.

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Endothelial Cell Culture Protocol

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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 (GlutaMAX^TM; 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.

Tomita Y., Palethorpe H.M., Smith E., Nakhjavani M., Townsend A.R., Price T.J., Yool A.J, & Hardingham J.E. (2019). Bumetanide-Derived Aquaporin 1 Inhibitors, AqB013 and AqB050 Inhibit Tube Formation of Endothelial Cells through Induction of Apoptosis and Impaired Migration In Vitro. International Journal of Molecular Sciences, 20(8), 1818.

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Cytotoxicity Evaluation of Bioresorbable Scaffolds

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Human umbilical vein endothelial cells (HUVECs; ATCC, Manassas, VA) were expanded in growth media consisting of Endothelial Cell Growth Kit-VEGF (ATCC^® PCS-100-041) in Vascular Cell Basal Medium (ATCC^® PCS-100-030) under the standard culture condition (37 °C with 5% CO2 in a humid environment) to 80% confluency before passaging. HUVECs at passages 5-7 were used.
The effect of the iodixanol introduction on the cytotoxicity of BRS was evaluated by means of an indirect cytotoxicity assay following ISO 10993-5: 2009. Specifically, the BRS were first sterilized by Ethylene Oxide and 0.5 mL of growth media was incubated with BRS for 24 hours at 37 °C with agitation. Next, the liquid extracts of different BRS were collected and added to HUVECs at sub-confluence in 24-well pates. After incubation for 24 hours, cells were incubated with growth media containing 0.5 mg/mL MTT (Thermo Fisher Scientific, San Jose, CA) for 3 hours. After replacing the MTT solution with DMSO, the plate was shaken for 15 min to solubilize the formazan crystals, and the absorbance of each well was measured at 570 nm using a microplate reader Cytation 5 (BioTek Instruments). The number of viable cells per well was calculated against a standard curve prepared by plating various concentrations of isolated cells, as determined by hemocytometer, in triplicate in the culture plates.

Ding Y., Fu R., Collins C.P., Sun C, & Ameer G.A. (2022). 3D-Printed Radiopaque Bioresorbable Stents to Improve Device Visualization. Advanced healthcare materials, 11(23), e2201955.

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Culturing Primary Human Airway and Vascular Cells

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Primary Human Bronchial Epithelial Cells (HBECs, ATCC) cells were cultured in Airway Epithelial Cell Basal Medium (Cat. No. PCS-300-030, ATCC) supplemented with Bronchial Epithelial Cell Growth Kit (Cat. No. PCS-300-040, ATCC). Primary Human Pulmonary Artery Endothelial Cells (HPAECs, ATCC) were grown in Vascular Cell Basal Medium (Cat. No. PCS-100-030, ATCC), supplemented with Endothelial Cell Growth Kit-VEGF (Cat. No. PCS-100-041, ATCC) The cells were maintained in standard conditions at 37 °C, 5% CO₂, and 95% humidity.

Targosz-Korecka M., Kubisiak A., Kloska D., Kopacz A., Grochot-Przeczek A, & Szymonski M. (2021). Endothelial glycocalyx shields the interaction of SARS-CoV-2 spike protein with ACE2 receptors. Scientific Reports, 11, 12157.

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Insulin Resistance in Endothelial Cells

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Endothelial cells (EC) from human aorta (TeloHAEC cell line, CRL-4052) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and grown as a monolayer, at 37 °C under a humidified atmosphere with 5% CO₂, in Vascular Cell Basal Medium (PCS-100-030, ATCC, Manassas, VA, USA) supplemented with Endothelial Cell Growth kit-VEGF (PCS-100-041, ATCC, Manassas, VA, USA). To induce insulin-resistance (IR), EC were exposed to palmitic acid (PA) up to 1.5 mM for 24 and 48 h. To confirm the IR assessment, the 48 h treatment with PA was followed by incubation with 100 nM recombinant human insulin (91077C, Merck KGaA, Darmstadt, Germany) for 30 min, as previously described [26 (link)]. δ-Valerobetaine (δVB) synthesis and purification were carried out as previously described [25 (link)]. δVB was dissolved in Hanks’ balanced salt solution (HBSS)-10 mM Hepes and treatments were performed for a maximum time of 72 h with betaine concentrations from 0.1 up to 1 mM. For combined treatments (δVB+PA), EC were pre-treated for 16 h with 0.5 mM δVB, followed by removal of δVB and exposure to PA (0.5 mM) for 48 h. Control cells (Ctr) were maintained in a complete culture medium with the corresponding volume of HBSS-10 mM Hepes.

Martino E., Balestrieri A., Anastasio C., Maione M., Mele L., Cautela D., Campanile G., Balestrieri M.L, & D’Onofrio N. (2022). SIRT3 Modulates Endothelial Mitochondrial Redox State during Insulin Resistance. Antioxidants, 11(8), 1611.

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Culturing Ovarian Cancer and Normal Cells

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Human ovarian cancer cell line A2780/CP70 and human normal ovarian cells IOSE-364 were cultured in RPMI 1640 medium (Sigma, St. Louis, MO, USA) supplemented with 10% US-qualified fetal bovine serum (Invitrogen, Grand Island, NY, USA) at 37 °C with 5% CO₂. HUVECs were cultured in vascular basal medium supplemented with endothelial cell growth kit-VEGF (ATCC, Manassas, VA, USA) at 37 °C with 5% CO₂.

Zhang Y., Chen S., Wei C., Rankin G.O., Rojanasakul Y., Ren N., Ye X, & Chen Y.C. (2017). Dietary Compound Proanthocyanidins from Chinese bayberry (Myrica rubra Sieb. et Zucc.) leaves inhibit angiogenesis and regulate cell cycle of cisplatin-resistant ovarian cancer cells via targeting Akt pathway. Journal of functional foods, 40, 573-581.

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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 described^{19 (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.

Kudelka M.R., Antonopoulos A., Wang Y., Duong D.M., Song X., Seyfried N.T., Dell A., Haslam S.M., Cummings R.D, & Ju T. (2015). Cellular O-Glycome Reporter/Amplification to Explore O-Glycans of Living Cells. Nature methods, 13(1), 81-86.

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