VR1814, a clinical isolate of CMV that was adapted for growth in HUVEC [87 (link)], was a generous gift (at passage 23) from Dr. Maria Grazia Revello. Virus was propagated in HUVEC (Lonza), and viral stocks at passages 27–29 were prepared from supernatant virus [126 , 127 (link)]. The titers of infectious virus were determined by a rapid method of immunological detection and quantification of CMV IE proteins that has been shown to correlate with the conventional plaque assay [126 , 128 (link)]. Neonatal human dermal fibroblasts (NHDF-Neo) (Lonza) were used as the indicator monolayer for the assay, and virus titers are expressed as IU [126 ]. HCoEpiC isolated from human fetal colonic tissue were purchased from ScienCell at passage 1. HCoEpiC were cultured in colonic epithelial cell medium (CoEpiCM, ScienCell), and all experiments were performed at passage 2 or 3. The purity of colonic epithelial cells was verified at each passage by immunofluorescence staining of formaldehyde-fixed cytospin preparations with a rabbit monoclonal antibody to human cytokeratin 19 (Abcam) (S3 Fig ).
Coepicm
Manufactured by ScienCell
Sourced in Japan, United States
CoEpiCM is a culture medium designed for the growth and maintenance of co-cultured epithelial cells. It provides a balanced formulation of nutrients, growth factors, and other essential components to support the co-culture of epithelial cell types.
Automatically generated - may contain errors
Lab products found in correlation
Hcoepic, by ScienCell (2 mentions)
Human umbilical vein endothelial cells (huvec), by Lonza (1 mentions)
Nhdf neo, by Lonza (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Bronchial epithelial cell growth basal medium, by Lonza (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
Apo transferrin, by Merck Group (1 mentions)
Panc 1, by American Type Culture Collection (1 mentions)
Insulin, by Merck Group (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Sodium selenite, by Merck Group (1 mentions)
Suit 2, by Health Science Research Resources Bank (1 mentions)
Bay 11 7085, by Merck Group (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Tnf α, by Merck Group (1 mentions)
5 protocols using coepicm
1
CMV Isolation and Quantification in HUVEC and HCoEpiC
2
Cell Culture Conditions for Various Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H226, HCT116, DLD1, DMS114 and DMS273 cells were cultured in Roswell Park Memorial Institute medium (RPMI medium 1640 (1×), Gibco 11875-093), supplemented with 10% (v/v) fetal bovine serum, 1% (v/v) Penicillin-Streptomycin (Gibco, 15410-122). A549 and HT29 cells were cultured in Dulbecco’s Modified Eagle medium (DMEM medium (1×), Gibco 11885-084), supplemented with 10% (v/v) fetal bovine serum, 1% (v/v) Penicillin-Streptomycin (Gibco, 15410-122). NHBE cells were cultured in Bronchial Epithelial Cell Basal medium (BEBM, Lonza CC-3170), supplemented with Bronchial Epithelial Cell Growth Medium SingleQuots Supplements and Growth Factors (BEGM, Lonza, CC-4175). HCoEpic cells were cultured in Colonic Epithelial Cell Medium (CoEpiCM, ScienCell, #2951), supplemented with Colonic Epithelial Cell Growth Suppllement (CoEpiCGS, ScienCell, #2952). All cells were cultured in a humidified incubator under 5% CO2 in 95% air.
3
Culturing Human Colonic Epithelial and Kidney Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human colonic epithelial cells (HCoEpiCs) were purchased from ScienCell Research Laboratories and cultured in colonic epithelial cell medium (CoEpiCM, ScienCell Research Laboratories). Human embryonal kidney 293T cells were cultured in DMEM containing 10% FBS. The cells were cultured at 37°C in 5% CO2.
4
Culturing Human Cancer and Normal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human cancer cell lines were grown in McCoy’s 5A Medium (HCT116 [ATCC]), Roswell Park Memorial Institute (RPMI) 1640 (MKN45 [Japanese Cancer Research Resources Bank, Tsukuba, Japan], PANC-1 [ATCC], OE33 [DS Pharma Biomedical Co., Ltd., Osaka, Japan]) or high-glucose Dulbecco’s modified Eagle’s medium (DMEM) (SUIT-2 [Health Science Research Resources Bank, Osaka, Japan]) supplemented with 10% (vol/vol) fetal bovine serum (FBS), 2 mM l -glutamine, 50 U/ml penicillin, and 50 µg/ml streptomycin in a humidified atmosphere containing 5% CO2. Human colorectal epithelial primary (CoEpiC; ScienCell Research Laboratories, Inc., CA, USA), Het1A (non-tumorous esophagus cells, ATCC) and HCEC-1CT (Summit Pharmaceuticals International Corporation, Tokyo, Japan) cells were grown in colonic epithelial cell medium (CoEpiCM; ScienCell), bronchial epithelial cell growth basal medium (Lonza, Basel, Switzerland) and ColoUp medium (DMEM/Medium 199 Earle’s, 4 + 1 (Biochrom Cat# F0435 and Cat# FG0615) containing 4 mM GlutaMAXTM-1 (100×), (Gibco, Cat# 35050-038) 2% cosmic calf serum (Hyclone, Cat# SH30087), 20 ng/ml EGF (Sigma Aldrich, Cat# E9644), 10 μg/ml Insulin (Sigma Aldrich, Cat# I9278), 2 μg/ml Apo-Transferrin (Sigma Aldrich, Cat# T2036), 5 nM sodium-selenite (Sigma Aldrich, Cat# S5261), and 1 μg/ml hydrocortisone (Sigma Aldrich, Cat# H0396)), respectively.
5
Inducing Ferroptosis in HCoEpiC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HCoEpiC cell (human normal colonic epithelial cell) was cultured in colonic epithelial cell medium (CoEpiCM, ScienCell Research Laboratory, CA, USA) containing 10% fetal bovine serum and other supplements according to the manufacturer's instructions (ScienCell Research Laboratory). To induce ferroptosis, cells were seeded on 12-well plates and treated with RSL3 (Selleck, 20 μm ) for 8 hours after plating. For the ER stress suppression experiment, 1 μm GSK 414 was added to the medium 30 minutes before RSL3 challenge. For the TNF-α treatment experiment, the cells were administrated with TNF-α (PeproTech, Rocky Hill, NJ, USA, 40 ng/ml) for 1 hour and the NF-κB inhibitor, BAY 11-7085 (Merck, 10 μm ), was added to some wells 15 minutes before the TNF-α was added.
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!