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Egm 2 singlequot kit

Manufactured by Lonza
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Sourced in United States, Switzerland

The EGM-2 SingleQuot Kit is a specialized laboratory product designed for cell culture applications. It provides a standardized and pre-formulated set of growth supplements that are essential for the optimal growth and maintenance of endothelial cells in vitro. The kit contains a balanced mixture of factors, including growth factors, hormones, and other critical components, to support the specific needs of endothelial cell cultures.

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Lab products found in correlation

Ebm 2 basal medium, by Lonza (6 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions) Huvecs, by Lonza (4 mentions) Cell culture grade, by Merck Group (3 mentions) Egf 236 eg, by R&D Systems (3 mentions) Tak 165, by Bio-Techne (3 mentions) Dispase 2, by Roche (3 mentions) Collagenase 2, by Worthington (3 mentions) Human umbilical vein endothelial cells (huvec), by Lonza (3 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions) Ebm 2, by Lonza (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Hmscs, by RoosterBio (2 mentions) Ascorbic acid, by Merck Group (2 mentions) Dexamethasone (dex), by Merck Group (2 mentions) β glycerophosphate, by Merck Group (2 mentions) D glucose and dnase 1, by Merck Group (2 mentions) Gi254023x, by Bio-Techne (2 mentions) Egm singlequot kit, by Lonza (2 mentions) Trizol, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

EGM-2 SingleQuots EGM™ SingleQuots™ Kit EGM SingleQuot Kit EGM SingleQuots EGM-2 kit SingleQuots kit SingleQuot Kit EGM-2 SingleQuots

30 protocols using egm 2 singlequot kit

1

Neuroblastoma BE(2)-C and HUVEC Cell Culture

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Neuroblastoma BE(2)-C cells were cultured in RPMI 1640 (Mediatech, Cat No. 10-040-CV) supplemented with 10% fetal bovine serum (Sigma-Aldrich, Cat No. F2442) at 37°C and humidified 5% CO2. HUVECs obtained from Dr. M. Freeman (Vanderbilt University Medical Center) were cultured in EMM-2 supplemented with Growth Factors (EGM-2 SingleQuot kit, Lonza, Cat No. CC-4176) at 37°C and humidified 5% CO2. The miR-CON, pCMV-miR-145 plasmid was obtained from Dr. Yingjie Yu (Wayne State University School of Medicine). Cisplatin (CDDP) was purchase from Sigma-Aldrich (Cat No 15663-27-1), and Vincristine was purchased from Sigma-Aldrich (Cat No. V8879). The primary antibodies used include LC3-1&II (MBL, PD014), ꞵ-actin antibody was from Sigma-Aldrich (1:5000, A2066) and Bafilomycin A1 (BafA1) was purchased from LC Laboratories (B-1080).
Kim K.W., Qiao J., Kim J.Y., Park K, & Chung D.H. (2020). Overexpression of microRNA-145 inhibits tumorigenesis through autophagy in chemotherapy and radiation resistant neuroblastoma cells. Oncoscience, 7(1-2), 1-9.
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2

HUVEC Culture Protocol for Cell Stretching

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Human umbilical vein endothelial cells (HUVECs; Lonza) were cultured in the endothelial basal medium (EBM-2, Lonza) supplemented with fetal bovine serum (FBS) and soluble factors (EGM-2 SingleQuot Kit, Lonza) including hEGF, VEGF, hFGF-B, R3-IGF-1, heparin, ascorbic acid, hydrocortisone, gentamicin, and amphotericin. Cells were maintained on tissue culture plates pre-coated with 0.1% gelatin (w/v; Sigma-Aldrich). After reaching 70% – 90% confluence, HUVECs were trypsinized (0.025% trypsin/EDTA, Lonza) and passaged with a seeding density of 5 × 103 cells cm−2. Before cell stretching assays, single HUVECs were plated at a density of 3 × 103 cells cm−2 on the PDMS basal membrane. Cells were cultured at 37°C and 5% CO2 and allowed to attach to the PDMS membrane overnight before downstream assays. Only early passages of HUVECs (passage 3 – 6) were used in this study.
Shao Y., Mann J.M., Chen W, & Fu J. (2014). Global architecture of F-actin cytoskeleton regulates cell shape-dependent endothelial mechanotransduction. Integrative biology : quantitative biosciences from nano to macro, 6(3), 300-311.
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3

Culturing HUVEC and HL-60 Cells

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Human umbilical vein endothelial cells (HUVECs) obtained from Lonza (Walkersville, MD, USA) were cultured in EBM-2 basal medium supplemented with EGM-2 SingleQuot Kit (Lonza), which contained 10% FBS,38 (link) according to the manufacturer’s instruction, and used for experiments at passage 4–6. The HL-60 human promyelocytic leukemia cell line (ATCC, Manassas, VA, USA) was cultured and differentiated into a neutrophil-like lineage as described earlier.30 (link),41 (link) All cell cultures were maintained at 37°C in 5% CO2.
Yakovlev S., Cao C., Galisteo R., Zhang L., Strickland D.K, & Medved L. (2019). Fibrin-VLDL receptor-dependent pathway promotes leukocyte transmigration by inhibiting Src kinase Fyn and is a target for fibrin β15–42 peptide. Thrombosis and haemostasis, 119(11), 1816-1826.
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4

HAEC Exposure to TGRL Lipolysis

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Human aortic endothelial cells (HAEC) (passage 6, Cascade Biologics, Portland, OR) were cultured in EBM-2 basal media (cc-3156) supplemented with EGM-2 SingleQuot Kit (cc-4176) (Lonza, Walkersville MD) under an atmosphere of 5% CO2: 95% air at 37°C. Cells were exposed for 1, 2, and 3 h to the following conditions, media or TGRL lipolysis products (TGRL (150 mg/dL = 1.5 mg/mL) + lipoprotein lipase LpL (2 U/mL). The final concentration of TGRL lipolysis products were diluted in media and pre-incubated for 30 minutes at 37°C prior to application.
Yahiatène I., Aung H.H., Wilson D.W, & Rutledge J.C. (2014). Single-molecule quantification of lipotoxic expression of activating transcription factor 3. Physical chemistry chemical physics : PCCP, 16(39), 21595-21601.
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5

U87MG and HUVEC Cell Culture

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The human GB cell line U87MG was obtained from the ATCC (LGC Promochem, Molsheim, France) and expanded in DMEM-high glucose medium (DMEM-HG, Ozyme) containing 10% FBS (Fisher Scientific, Illkirch, France) and 1% antibiotics (Sigma-Aldrich). Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza (Verviers, Belgium). Cells were cultured, according to the supplier’s instructions, in endothelial cell growth medium-2 (EGM-2), corresponding to endothelial basal medium-2 (EBM-2) containing the supplements and growth factors of the EGM-2 SingleQuot™ kit (Lonza). U87MG cells and HUVECs were maintained under an atmosphere containing 5% CO2 (37 °C), in a humidified incubator, until they reached 80% confluence.
Clavreul A., Roger E., Pourbaghi-Masouleh M., Lemaire L., Tétaud C, & Menei P. (2018). Development and characterization of sorafenib-loaded lipid nanocapsules for the treatment of glioblastoma. Drug Delivery, 25(1), 1756-1765.
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6

Culturing Neuroblastoma and HUVEC Cells

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All neuroblastoma cell lines BE(2)-C, SK-N-AS, BE-M17, SK-N-SH, and SH-SY5Y were purchased from the American Type Culture Collection (ATCC, Manassas, VA). Cells were maintained in RPMI 1640 with 10% Fetal Bovine Serum (FBS) at 37°C in a humidified atmosphere consisting of 5% CO2 and 95% air. HUVECs obtained from Dr. M Freeman (Vanderbilt University Medical Center, Nashville, TN) were cultured in EMM-2 supplemented with growth factors (EGM-2 Single- Quot kit, Lonza, Walkersville, MD, USA) at 37°C and humidified 5% CO2.
Qiao J., Rellinger E.J., Kim K.W., Powers C.M., Lee S., Correa H, & Chung D.H. (2019). Identification of α-N-catenin as a novel tumor suppressor in neuroblastoma. Oncotarget, 10(49), 5028-5040.
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7

HUVEC Culture in EGM-2 Medium

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Human umbilical vein endothelial cells (HUVEC; Lonza, C2519A, Basel, Switzerland) were cultured in endothelial basal medium-2 containing EGM-2 SingleQuot Kit (Lonza, CC-3162, Basel, Switzerland). The medium was renewed every 48 h. Cells were cultured at 37 °C and in a CO2 air atmosphere with a humidity of 5% (v/v).
Kim S.Y., Kim Y.J., Kim S., Momeni M., Lee A., Treanor A., Kim S., Kim R.H, & Park N.H. (2023). GV1001 Inhibits the Severity of the Ligature-Induced Periodontitis and the Vascular Lipid Deposition Associated with the Periodontitis in Mice. International Journal of Molecular Sciences, 24(16), 12566.
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8

Investigating Endothelial Cell Responses

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Human aortic endothelial cells (HAECs), endothelial basal medium-2 (EBM-2), and EGM-2 SingleQuot kit supplements and growth factors were purchased from Lonza (Allendale, NJ). Fetal bovine serum (FBS), cell culture supplements, calcein-AM, and RPMI 1640 were from Invitrogen (Carlsbad, CA); cell titer blue from Promega (Madison, WI); protein assay kits were from Bio-Rad (Hercules, CA); fatty acid-free BSA from Fisher Scientific (Hampton, NH); palmitate, protease and phosphatase inhibitor cocktails, malvidin-3-glucoside, cyanidin-3-glucoside, hippuric acid, hydroxyhippuric acid, 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF), 2′,7′-dichlorofluorescein diacetate, and other general chemicals were obtained from Sigma–Aldrich (St. Louis, MO). Human monocytic THP1 cells were from American Type Culture Collection (Manassas, VA). RNeasy plus mini kit, QuantiTech reverse transcription kit, SYBR qPCR (quantitative PCR) green master mix, and primers were purchased from Qiagen (Valencia, CA).
Bharat D., Cavalcanti R.R., Petersen C., Begaye N., Cutler B.R., Costa M.M., Ramos R.K., Ferreira M.R., Li Y., Bharath L.P., Toolson E., Sebahar P., Looper R.E., Jalili T., Rajasekaran N.S., Jia Z., Symons J.D, & Babu P.V. (2017). Blueberry Metabolites Attenuate Lipotoxicity-Induced Endothelial Dysfunction. Molecular nutrition & food research, 62(2), 10.1002/mnfr.201700601.
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9

Primary Pulmonary Endothelial Cell Culture

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Human primary pulmonary artery endothelial cells (HPAEC) were purchased from ATCC (ATCC) and maintained in endothelial cell growth medium-2 (EGM-2, Lonza) supplemented with EGM-2 SingleQuot Kit (Lonza). HPAEC were cultured in multi-well cell culture plates and flasks coated with 0.1% gelatin. All experiments were performed in cells cultured for ≤7 passages. DMOG (Tocris) was dissolved in dimethyl sulfoxide (DMSO). Trimethyl citrate (Sigma), methyl pyruvate (Thermo Scientific™), methyl aspartate (Santa Cruz) and nicotinamide riboside (Cayman) were used for metabolic repletion experiments.
Tiwari R., Bommi P.V., Gao P., Schipma M.J., Zhou Y., Quaggin S.E., Chandel N.S, & Kapitsinou P.P. (2022). Chemical inhibition of oxygen-sensing prolyl hydroxylases impairs angiogenic competence of human vascular endothelium through metabolic reprogramming. iScience, 25(10), 105086.
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10

Culturing Primary Pulmonary Arterial Endothelial Cells

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Primary PAECs (Lonza, Frederick, MD, USA) were cultured for a maximum of 6 passages on cell culture flasks coated with type I collagen (50 μg/mL in 0.02 N acetic acid; Corning, Corning, NY, USA) in endothelial basal medium-2 (EBM™-2, Lonza, Frederick, MD, USA) supplemented with growth factors and 2% serum (EGM™-2 SingleQuot kit, Lonza, Frederick, MD, USA). Failed donor control (CTRL) and IPAH endothelial cells (ECs) obtained from the Pulmonary Hypertension Breakthrough Initiative (PHBI) (see Supplementary Table S3 donor information) were plated on 0.1% gelatin (Sigma, St. Louis, MO, USA). In experiments studying exogenous DLL4 effects, cells were seeded on plates pre-coated with recombinant DLL4 (0.5 μg/mL in PBS overnight, R&D Systems; Minneapolis, MN, USA). Cells were maintained at 37 °C in a humidified incubator with 5% CO2 and 95% air.
Awad K.S., Wang S., Dougherty E.J., Keshavarz A., Demirkale C.Y., Yu Z.X., Miller L., Elinoff J.M, & Danner R.L. (2024). BMPR2 Loss Activates AKT by Disrupting DLL4/NOTCH1 and PPARγ Signaling in Pulmonary Arterial Hypertension. International Journal of Molecular Sciences, 25(10), 5403.
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