Ebm 2 culture medium

Manufactured by Lonza

Sourced in United States, Switzerland

The EBM-2 culture medium is a sterile liquid product used for the in vitro cultivation of mammalian cells. It provides the necessary nutrients and growth factors to support the growth and proliferation of various cell types in a controlled laboratory environment.

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Lab products found in correlation

Egm 2 bulletkit, by Lonza (3 mentions) Huvecs, by Lonza (3 mentions) Reverse transcription kit, by Thermo Fisher Scientific (3 mentions) Blebbistatin, by LGC (1 mentions) Mcdb 131 medium, by Thermo Fisher Scientific (1 mentions) X tremegene hp dna transfection reagent, by Roche (1 mentions) Nocodazole, by Merck Group (1 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Calyculin a, by Merck Group (1 mentions) Blebbistatin, by Merck Group (1 mentions) Huvecs, by Corning (1 mentions) Y 27632, by Merck Group (1 mentions) Incucyte live cell imaging system, by Sartorius (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Endothelial mitogen, by Biomedical Technologies (1 mentions) C2519as, by Lonza (1 mentions) Egm 2 singlequot, by Lonza (1 mentions)

8 protocols using ebm 2 culture medium

Isolation and Expansion of hASCs and HUVECs

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Humans ASCs were a kind gift of Dr. Jeffrey Gimble. The hASCs were isolated under an Institutional Review Board approved protocol according to published methods [24 ]. Cells from pooled donors were used at passages 3 – 5.
A vial of pooled Human Umbilical Vein Endothelial Cells (HUVECs) (Lonza, USA) were expanded in tissue culture flasks up to Passage 5 using Endothelial Basal Medium 2 (EBM-2) culture medium (Lonza, USA) supplemented with the Endothelial Growth Medium-2 (EGM-2) Bullet Kit (Hydrocortisone, human fibroblast growth factor (hFGF)-2, vascular endothelial growth factor (VEGF), R3-insulin-lke growth factor (IGF)-1, ascorbic acid, human epidermal growth factor (hEGF), Gentamicin & Amphotericin (GA-1000), and Heparin) and 2% FBS. HUVECs were used for experimentation at Passage 6.

Cook C.A., Hahn K.C., Morrissette-McAlmon J.B, & Grayson W.L. (2015). Oxygen Delivery from Hyperbarically Loaded Microtanks Extends Cell Viability in Anoxic Environments. Biomaterials, 52, 376-384.

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Endothelial Cell Culture and Transfection

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Human dermal microvascular endothelial cells (HMECs; Ades et al., 1992 (link)) were grown in MCDB 131 medium (Gibco) supplemented with 10% FBS, 0.003 mg/ml human EGF, 0.001 mg/ml hydrocortisone, and l-glutamine. Primary human pulmonary arterial endothelial cells were grown in EBM-2 culture medium (Lonza) supplemented with 15% FBS and EGM-2 bullet kit (Lonza) and were used at passages 2–6. All cell lines were maintained at 37°C and 5% CO₂.
Endothelial cells were plated on glass-bottom coverslips coated with 0.2% gelatin and transfected at 70–80% confluency using X-tremeGENE HP DNA transfection reagent according to manufacturer’s protocol (Roche). For adenoviral infection, endothelial cells were exposed to the adenoviral particles overnight and were used for live-cell imaging at 24–72 h after infection. The procedure was handled according to National Institutes of Health safety guidelines for materials containing BSL-2 organisms. Cells were treated with 20 µM cRO or 10 µM blebbistatin (Toronto Research Chemicals) for 10 min before experiments.

Daneshjou N., Sieracki N., van Nieuw Amerongen G.P., Conway D.E., Schwartz M.A., Komarova Y.A, & Malik A.B. (2015). Rac1 functions as a reversible tension modulator to stabilize VE-cadherin trans-interaction. The Journal of Cell Biology, 208(1), 23-32.

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HUVEC Culture Conditions and Treatments

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Human Umbilical Vein Endothelial Cells (HUVECs) were purchased from Corning Inc. HUVECs were cultured in endothelial basal medium-2 (EBM-2) culture medium (Lonza Inc.) with 2% fetal bovine serum. Cells were grown at 37°C and 5% CO2 on tissue culture dishes in a humidified incubator with media changes every two days. Cells were passaged using 0.25% Trypsin (Invitrogen), and passages 2-7 were used in the experiments. DAPT, Y-27632, blebbistatin, nocodazole and calyculin A were purchased from Sigma Aldrich. Jag1 peptide (188-204) was acquired from Anaspec. HUVECs were treated with DAPT (20 μM), Jag1 peptide (20 μM), blebbistatin (20 μM), nocodazole (1 μM), and calyculin A (5 nM).

Wang S., Sun J., Xiao Y., Lu Y., Zhang D.D, & Wong P.K. (2017). Intercellular Tension Negatively Regulates Angiogenic Sprouting of Endothelial Tip Cells via Notch1-Dll4 Signaling. Advanced biosystems, 1(1-2), 1600019.

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Placenta-Derived Exosomes Promote Angiogenesis

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Example 8

Human vascular endothelial cells (HUVECs) at passage 3 or 4 were plated in 48-well microtiter dishes at a density of 28,000 cells per well. Cells were maintained in 0.5 mL of either PBD or EBM-2 culture medium (Lonza) on GRF-Matrigel for 3 days. HUVECs were then incubated with different concentrations of placenta-derived adherent cell exosomes isolated from either serum-containing (“growth exosomes”) or serum-free (“SF exosomes”) placenta-derived adherent cell cultures. HUVEC growth patterns, including node formation and tube segment length (surrogates for angiogenic potential) were monitored every two hours for 18-19 hours total. HUVEC growth parameters were determined using IncuCyte live cell imaging system (Essen BioScience) and quantified using ImageJ.

US11173182B1. Placenta-derived adherent cell exosomes and uses thereof (2021-11-16). Celularity Inc. [US]. Inventors: Eric Law [US], Andrew Morschauser [US], Aleksander Francki [US], Jennifer Paredes [US], Kathy Karasiewicz-Mendez [US], Allan Reduta [US], Vladimir Jankovic [US], Ivana Djuretic [US], Robert J. Hariri [US].

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Culturing Endothelial Cells from Different Sources

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Pooled human umbilical vein endothelial cells (HUVECs) from different donors (LONZA, Walkersville, MD, USA) were cultured in EBM-2 culture medium supplemented with EGM-2 bullet kit (LONZA) on gelatin-coated tissue dishes. HUVECs up to passage 6 were used for experiments. COS-7 cells, transformed African green monkey kidney fibroblast cells (ATCC, Manassas, VA, USA), and HEK293T cells (a gift from Dr. Yuanfei Wu, Yale University, New Haven, CT, USA) were cultured in Dulbecco's modified Eagle's medium (DMEM) (Invitrogen, Grand Island, NY, USA) supplemented with 10% fetal bovine serum and penicillin/streptomycin (Invitrogen). Primary mouse lung microvascular endothelial cells were isolated from mice lung by the method previously described [63] (link), [64] (link) and cultured in DMEM supplemented with 20% fetal bovine serum, penicillin/streptomycin, non-essential amino acids (Invitrogen) and 20 µg/ml endothelial mitogen (Biomedical Technologies, Inc., Stoughton, MA, USA).

Sakurai T., Woolls M.J., Jin S.W., Murakami M, & Simons M. (2014). Inter-Cellular Exchange of Cellular Components via VE-Cadherin-Dependent Trans-Endocytosis. PLoS ONE, 9(3), e90736.

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Culturing Human Umbilical Vein Endothelial Cells

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Human umbilical vein endothelial cells (C2519AS, Lonza, Basel, Switzerland) were cultured in plates coated with 0.1% gelatin in PBS in EBM-2 culture medium (CC-3156, Lonza, Basel, Switzerland) supplemented with EGM-2 SingleQuots (CC-4176, Lonza, Basel, Switzerland), and were between passages 3 and 4. The cells were incubated at 37 °C in a humidified 5% CO₂ environment.

Peeters J.A., Peters H.A., Videler A.J., Hamming J.F., Schepers A, & Quax P.H. (2023). Exploring the Effects of Human Bone Marrow-Derived Mononuclear Cells on Angiogenesis In Vitro. International Journal of Molecular Sciences, 24(18), 13822.

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Cell Culture of Human Ovarian and Lung Cells

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Human ovarian cancer cell lines, SKOV-3 and OVCAR-3, were purchased from American Type Tissue Culture Collection (ATCC, Manassas, VA, USA). Cells were cultured at 37 °C in 5% CO₂ humidified atmosphere in either McCoy’s 5A modified medium with 10% FBS and 1% antibiotics or RPMI-1640 medium with 20% FBS, 1% antibiotics, and 0.01 mg/mL bovine insulin, respectively. Unless otherwise stated, all reagents were purchased from Merck KGaA, Darmstadt, Germany.
Human lung microvascular endothelial cells (HMVEC) were purchased from Lonza and were cultured in EBM-2 culture medium supplemented with EGM-2 MV SingleQuots (Lonza, Basel, Switzerland).

Kosowska A., Garczorz W., Kłych-Ratuszny A., Aghdam M.R., Kimsa-Furdzik M., Simka-Lampa K, & Francuz T. (2020). Sitagliptin Modulates the Response of Ovarian Cancer Cells to Chemotherapeutic Agents. International Journal of Molecular Sciences, 21(23), 8976.

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Endothelial Cell Isolation and Characterization

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EBM-2 culture medium (Lonza, USA), ficoll hypaque 1.077 (Haoyang, Tianjin, China), lipopolysaccharide (Solarbio, Beijing, China), CCK-8 kit (Dojindo Laboratories, Japan), FITC-coupled annexin-V apoptosis detection kit (BD Biosciences, USA), cell cycle and apoptosis kit (Beyotime Biotechnology, Shanghai, China), FITC-UEA-I/ DiI-acLDL (Proteintech, USA), PCDH-CMV-EF1-copGFP-Sirt1 (LabGene, Guangzhou, China), SRT1720/EX527 (Sigma-Aldrich, USA), reverse transcription kit (Thermo Fisher, USA), rabbit monoclonal antibodies to Sirt1/β-actin (ab189494/ ab115777, Abcam, UK), rabbit monoclonal antibodies to p53/NF-κB/FOXO3a (32532S/8242S/12829S, Cell Signaling, USA), HRP conjugated AffiniPure goat anti-rabbit (BA1055, BOSTER, Wuhan, China), PCR fluorescence quantification kit (Thermo Fisher, USA), Matrigel™ Matrix (BD Biosciences, USA), and CD34+ positive selection cocktail (EasySep, Stemcell, Canada) were used in the experiments.

Sun Y., Liu X., Sun D., Yao J., Zhan-ling D., Qian J., & Huang Q. (2021). Effects of Sirt1 on proliferation, migration, and apoptosis of endothelial progenitor cells in peripheral blood of SD rats with chronic obstructive pulmonary disease. Asian Pacific Journal of Tropical Biomedicine, 11(10), 429-429.

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