Colorectal cancer cell lines (WiDr, LS1034, and C2BBe1) purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) were used to perform in vitro studies. The three cell lines are from different colon localizations: WiDr from rectosigmoid location, C2BBe1 clone of the cell line Caco-2 (ATCC) from descending colon, and LS1034 resistant to chemotherapy concerning P-glycoprotein overexpression from ascending colon [9 (link)–13 (link)]. WiDr and C2BBe1 cell lines were maintained in Dulbecco's Modified Eagle's cell culture medium (Sigma) and LS1034 cell line was maintained in Roswell Park Memorial Institute (RPMI) medium. All cell lines were cultured in high (25 mM) and low (5 mM) glucose concentrations, supplemented with 10% fetal calf serum (Gibco) in 5% CO2 atmosphere, at 37°C. To perform hypoxia studies, cells were incubated at 37°C in 93% N2, 2% O2, and 5% CO2 using a controlled-environment cabinet (PlasLabs, USA). Each of the media was acquired without glucose in order to be supplemented with [U-13C]glucose for performing 13C NMR isotopomer analysis.
Dulbecco s modified eagle s cell culture medium
Manufactured by Merck Group
Dulbecco's Modified Eagle's cell culture medium is a widely used nutrient mixture for culturing a variety of mammalian cell lines. It provides essential nutrients, amino acids, and vitamins to support cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
C2bbe1, by American Type Culture Collection (2 mentions)
Ls1034, by American Type Culture Collection (2 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
C2bbe1, by Merck Group (1 mentions)
Ls1034, by Merck Group (1 mentions)
Caco 2, by American Type Culture Collection (1 mentions)
Sepmate pbmc isolation tube, by STEMCELL (1 mentions)
Countess 2 fl, by Thermo Fisher Scientific (1 mentions)
Sepmate conical tube, by STEMCELL (1 mentions)
Lymphoprep, by STEMCELL (1 mentions)
2 protocols using dulbecco s modified eagle s cell culture medium
1
Colon Cancer Cell Lines: In Vitro Studies
2
Isolation of Human PBMCs for ET-rich Thrombus
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To make ETs-rich thrombus analogs, human peripheral blood mononuclear cells (PBMCs) were added to mixture of human blood components to provide an abundant source of extracellular DNA. PBMCs were separated from human whole blood using the SepMate™ PBMC Isolation Tubes (STEMCELL Technologies) following the protocol provided by the company. Briefly, 12 mL human whole blood was diluted with Dulbecco's phosphate buffered saline (Cat #07905, STEMCELL Technologies) at 1:1 ratio, and then transferred into a 50 mL SepMate conical tube (Cat #85450, STEMCELL Technologies) pre-filled with 15 mL lymphoprep (Cat #07801, STEMCELL Technologies). The mixture was centrifuged at 1200 G for 10 mins at room temperature. The PBMCs layer was then harvested and subjected to repeated centrifugation at the same setting. The PBMCs were then washed and re-suspended in 1 ml Dulbecco's Modified Eagle's cell culture medium (Cat #D6546, Sigma-Aldrich). The cell counting and viability were assessed by an automated cell counter (Countess II FL, Thermo Fisher) using 0.4% Trypan blue solution. The resultant cell suspension had an average of 9.9x10 6 cells/mL with 80% viability.
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