Human embryonic kidney cells 293T (HEK293T), human bladder cancer cell line T24 and the human prostate cancer cell lines DU145 and PC3 were purchased from American Type Culture Collection (ATCC, Manassas, VA). Cell lines were maintained in DMEM or RPMI 1640 media as previously described 40 (link), 41 (link). Primary human umbilical vein endothelial cells (HUVEC, Cambrex Bio Science, Walkersville, MD, USA) were cultured in EBM-2 basal media supplemented with EGM-2 MV Kit (Lonza, Walkersville, MD) containing 2% FBS. HUVEC cells of passage 6 to 8 were used. The above cell lines were authenticated by Genetic Resources Core Facility at Johns Hopkins (Baltimore, MD). All cell lines were confirmed to be mycoplasma free.
Egm 2mv kit
Manufactured by Lonza
Sourced in United States
The EGM-2MV kit is a laboratory equipment product manufactured by Lonza. It is designed to support the growth and maintenance of endothelial cells in vitro. The kit includes a specialized endothelial growth medium and associated supplements.
Automatically generated - may contain errors
Lab products found in correlation
L glutamine, by Thermo Fisher Scientific (3 mentions)
Human umbilical vein endothelial cells (huvec), by Cambrex (2 mentions)
Hepes, by Thermo Fisher Scientific (2 mentions)
Endothelial basal medium 2, by Lonza (2 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions)
Ebm 2, by Lonza (2 mentions)
Du145, by American Type Culture Collection (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
In vivo jetpei, by Polyplus Transfection (1 mentions)
Verapamil hydrochloride vhcl, by Merck Group (1 mentions)
T75 culture flask, by Corning (1 mentions)
Ebm 2 medium, by Lonza (1 mentions)
Primary huvecs, by Cambrex (1 mentions)
Arp100, by Santa Cruz Biotechnology (1 mentions)
Gsk1120212, by Selleck Chemicals (1 mentions)
Alamarblue assay, by Thermo Fisher Scientific (1 mentions)
Pericyte medium pm, by ScienCell (1 mentions)
Fetal bovine serum (fbs), by ScienCell (1 mentions)
Penicillin streptomycin solution, by ScienCell (1 mentions)
Hdmvecs, by Lonza (1 mentions)
14 protocols using egm 2mv kit
1
Characterization of Human Cell Lines
2
Primary Adult HMVEC Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary adult HMVEC (Lonza Bioscience, Walkersville, MD) were cultured in Endothelial Basal Medium-2 (Lonza) supplemented with 5 % fetal bovine serum (FBS) and the growth factors contained within the EGM2-MV kit (Lonza), according to the manufacturer’s instructions. HFF and HaCaT (gifts from the laboratory of Dr. Jonathan Garlick, Tufts) were grown in DMEM supplemented with 10 % FBS (Atlanta Biologicals, Atlanta, GA), 1 % antibiotic–antimycotic, 1 % l -glutamine, and 10 mM HEPES (Life Technologies). HMVEC were used at P5–8, HFF were used at P14–22, and HaCaT were used at P40–46.
3
Comparative Analysis of Cancer and Benign Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human cervical cancer cell line HeLa (adenocarcinoma from ATCC, Manassas, VA) and benign human bladder cell line, UROtsa (a generous gift from Dr. Donald Sens at the University of North Dakota School of Medicine, Grand Forks, ND) were available for analysis. HeLa cells were maintained in RPMI 1640 media and UROtsa cells were maintained in DMEM media as previously described [12 (link)]. Primary human umbilical vein endothelial cell (HUVEC, Cambrex) was cultured in EBM-2 basal media supplemented with EGM-2 MV Kit (Lonza) containing 2% FBS. HUVEC cells of passage 6 to 8 were used. To ensure optimal siRNA delivery in xenograft tumors, in vivo-jetPEI (Polyplus-transfection Inc. NYC, NY), a linear polyethylenimine, was used in conjunction with siRNA [13 (link)].
4
Endothelial Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
7-ketocholesterol was purchased from Cayman Chemical (Ann Arbor, MI, USA). L-ergothioneine and L-ergothioneine-d9 (ET-d9; deuterated internal standard) were kindly provided by Tetrahedron (Paris, France). Tert-butyl hydroperoxide (TBHP), carbonyl cyanide 3-chlorophenylhydrazone (CCCP) and verapamil hydrochloride (VHCL), a nonspecific OCTN1 inhibitor, was purchased from Sigma Aldrich (St. Louis, MO, USA). Cells were cultured in Endothelial Basal Medium (EBM-2) (Lonza, Bend, OR, USA) with HyClone antimycotic/antibiotic solution (1×, Thermo Fisher Scientific, Waltham, MA, USA) and growth factors from the EGM-2MV kit (Lonza). The EGM-2MV kit contains gentamicin/amphotericin-B (GA), human epidermal growth factor (hEGF), ascorbic acid, vascular endothelial growth factor (VEGF), R3-insulin-like growth factor-1 (R3-IGF-1), hydrocortisone, human fibroblast growth factor-beta (hFGF-β) and foetal bovine serum (5% v/v).
5
Culturing Primary HMVEC in OrganoPlate
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary HMVEC (Human Microvascular Endothelial Cells) (Lonza CC-2543) were cultured in regular T75 culture flasks (Corning, 734–2705) with EBM2 medium (Lonza CC-3156) containing the EGM-2MV kit (Lonza-4147). HMVEC were passaged one time before being seeded into the OrganoPlate® 3-lane (MIMETAS 4003–400-B, Leiden, The Netherlands). Cell detachment was done with Trypsin/EDTA solution 0.25 mg/mL (Lonza CC-5012) and neutralized with Trypsin Neutralization solution TNS (Lonza CC-5002).
6
Bladder and Cervical Cancer Cell Lines
7
Human Retinal Endothelial Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human retinal endothelial cells (HREC; Cell Systems, Kirkland, Washington) were grown in complete endothelial base media (EBM-2 and EGM-2 MV Kit; Lonza, Walkersville, Maryland, USA). Cells from passages 3–7 were used in the experiments. To obtain optimum dosing range, viability was determined using Alamar blue assay (ThermoFisher Scientific).
8
Cultivation of Human Brain Cell Types
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human brain vascular pericytes (abbreviated as HBVP; obtained from ScienCell Research Laboratories, Carlsbad, CA, USA) were cultured on poly‐l ‐lysine‐coated dishes in Pericyte Medium (PM; ScienCell Research Laboratories) supplemented with 5% FBS (Sigma‐Aldrich, St. Louis, MO, USA) and penicillin/streptomycin solution (P/S; ScienCell Research Laboratories) and used between passage numbers 2 and 6. Human astrocytes (HA; ScienCell Research Laboratories) were grown on poly‐l ‐lysine‐coated dishes in astrocyte medium (AM) supplemented with 5% FBS and P/S (all from ScienCell Research Laboratories) and used between passage numbers 2 and 4. Human microvascular cerebral endothelial cells (hCMEC/D3, shortly D3) were cultured on rat‐tail collagen‐coated dishes in endothelial cell basal medium‐2 (EBM‐2) with EGM‐2MV kit including supplements and 2% FBS (all from Lonza, Basel, Switzerland) and used between passage numbers 30 and 40.
9
Culturing Vascular and Ocular Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were cultured at 37°C in 5% CO2. Primary HDMVECs were obtained from Lonza (Allendale, NJ) and cultured in EBM-2 base medium supplemented with the EGM-2 MV kit. HDMVECs were not used after 9 passages. Human 92.1 and OCM-1A UM cells were derived by and the generous gifts of Drs. Martine Jager (Laboratory of Ophthalmology, Leiden University) [19] (link) and June Kan-Mitchell (Biological Sciences, University of Texas at El Paso) [20] (link), respectively. Both cell lines were grown in RPMI 1640 medium (Life Technologies, Carlsbad, CA) supplemented with 10% FBS and antibiotics.
10
Culturing Primary Human Dermal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary adult human dermal microvascular endothelial cells (HMVEC, Lonza Bioscience, Walkersville, MD) were cultured in Endothelial Basal Medium-2 (Lonza) supplemented with 5% fetal bovine serum (FBS) and the growth factors contained within the EGM2-MV kit (Lonza), according to the manufacturer’s instructions. Primary adult human dermal fibroblasts (HDF, Lonza) were grown in Dulbecco’s Modified Eagle’s Medium (DMEM, Life Technologies, Grand Island, NY) supplemented with 10% FBS (Atlanta Biologicals, Atlanta, GA), 1% antibiotic-antimycotic, 1% L-glutamine, and 10mM HEPES (Life Technologies, Grand Island, NY). Primary adult human epidermal keratinocytes (NHEK, Lonza) were cultured as previously described [27 (link)]. HMVEC were used at P5-8, HDF were used at P3-8, and NHEK were used at P3-7.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!