U2OS, HCT116, and HEK293T cells were obtained from ATCC and cultured in D-MEM medium (U2OS and HEK293T) or RPMI 1640 (HCT116) supplemented with 10% fetal bovine serum, l -glutamine, penicillin, and streptomycin. OCI-AML3 cells were purchased from DSMZ GmbH and cultured in a D-MEM medium (PAN-Biotech) containing 20% FBS, l -glutamine, penicillin, and streptomycin. Cells were routinely tested for mycoplasma infection with a PCR-based method. For SILAC labeling, cells were cultured in media containing either l -arginine and l -lysine, l -arginine [13C6], and l -lysine [2H4] or l -arginine [13C615N4] and l -lysine [13C6-15N2] (Cambridge Isotope Laboratories)79 . All cells were cultured at 37 °C in a humidified incubator containing 5% CO2.
Dmem medium
Manufactured by American Type Culture Collection
Sourced in United States
DMEM medium is a commonly used cell culture medium that provides nutrients and growth factors necessary for the maintenance and proliferation of various cell types. It is a basal medium that can be further supplemented with additional components as required.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (11 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (10 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Dmem medium, by Thermo Fisher Scientific (4 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (3 mentions)
Hek293t, by American Type Culture Collection (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Rpmi 1640, by American Type Culture Collection (3 mentions)
Fetal calf serum (fcs), by American Type Culture Collection (2 mentions)
Rpmi 1640 medium, by American Type Culture Collection (2 mentions)
B16 f10 mouse, by American Type Culture Collection (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Penicillin, by American Type Culture Collection (2 mentions)
Non essential amino acids (neaa), by Corning (2 mentions)
Hepes, by Corning (2 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (2 mentions)
β mercaptoethanol, by Merck Group (2 mentions)
Mda mb 231, by American Type Culture Collection (2 mentions)
42 protocols using dmem medium
1
SILAC Labeling of Cell Lines
2
SW620 and LOVO Cell Line Culture
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SW620 (CCL-227; ATCC, American Type Culture Collection, Manassas, VA, USA) and LOVO (CCL-229; ATCC) cell lines were cultured with DMEM medium (cat. no. 12491-15; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS; cat. no. 10100-147; Thermo Fisher Scientific, Inc.) and 1% penicillin-streptomycin (cat. no. 15640055, Thermo Fisher Scientific, Inc.). The cell lines used in the present study were cultured at 37°C with 5% CO2.
Rg3 (cat. no. 64139; Sigma-Aldrich; Merck KGaA) and 5-FU (cat. no. 04541, Sigma-Aldrich; Merck KGaA) were dissolved in DMSO. For the cell experiments, the diluted culture solution of Rg3 or 5-FU was dissolved in DMSO to achieve the experimental concentration and was administered to the cells for 48 h. For animal experiments, PBS diluted Rg3 or 5-FU was dissolved in DMSO to the experimental concentration. The experiments were approved by the Ethics Committee of The Quanzhou First Hospital Affiliated to Fujian Medical University (Quanzhou, Fujian, China).
Rg3 (cat. no. 64139; Sigma-Aldrich; Merck KGaA) and 5-FU (cat. no. 04541, Sigma-Aldrich; Merck KGaA) were dissolved in DMSO. For the cell experiments, the diluted culture solution of Rg3 or 5-FU was dissolved in DMSO to achieve the experimental concentration and was administered to the cells for 48 h. For animal experiments, PBS diluted Rg3 or 5-FU was dissolved in DMSO to the experimental concentration. The experiments were approved by the Ethics Committee of The Quanzhou First Hospital Affiliated to Fujian Medical University (Quanzhou, Fujian, China).
3
Cell Culture Protocols for Cancer Research
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The human hepatoma Huh-7 cells and rat mammary tumor MRMT-1 cells were purchased from ATCC (Manassas, VA, USA). DMEM medium was purchased from ATCC (Manassas, VA, USA). Penicillin–streptomycin, 0.25% trypsin and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). Cells were cultured in DMEM supplemented with 10% FBS and 1% Penicillin–streptomycin in a humidified atmosphere at 37 °C with 5% CO2.
4
Tetracycline-Inducible TRPM7 Overexpression
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Tetracycline (Tet)-inducible HEK293-TREx cells, stably transfected with human TRPM7, were cultured in DMEM medium (Gibco) containing 10% fetal bovine serum (FBS, Gibco), zeocin (0.4 mg/ml, Invitrogen) and blasticidin (5 µg/ml, Invitrogen). Overexpression was induced adding 1 µg/ml Tetracycline (Gibco) to the culture medium. Patch-clamp experiments were performed 18–22 h post-Tetracycline induction. The normal rat kidney fibroblast cell line NRK-49F and epithelial cell line NRK-52E (ATCC) were maintained in DMEM medium (ATCC) supplemented with 5% fetal calf serum (ATCC). Cells were maintained at 37 °C under 5% CO2 condition.
5
Culturing and Treating Breast Cancer Cells
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MCF-7 cells (American Type Cell Culture, Manassas, VA) were cultured in RPMI 1640 medium (ATCC) with 10% Fetal Bovine Serum (ATCC). Caco-2 cells (a kind gift from Dr. Mohit Verma) were cultured with DMEM medium (ATCC) with 20% Fetal Bovine Serum and other additives (Final concentrations: 4.5 g/L Glucose, 10 mM HEPES, 44 mM Sodium Bicarbonate, 1 mM Sodium Pyruvate, 100 µM non-essential amino acids, 50 mg/L Gentamycin Sulfate, 100 U/L Penicillin, 100 U/L Streptomycin, 2 mM Glutamine, pH 7.2). MDA-MB-231 cells (ATCC) were also cultured in RPMI 1640 medium (ATCC) with 10% Fetal Bovine Serum. Paclitaxel (Selleck Chemicals, Houston, TX) prepared in DMSO (ATCC) was used to treat some of the tumors formed by single cells seeded on collagen I islands.
6
Culturing Brain Microvascular ECs and Glioma Cells
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Human and mouse brain microvascular ECs (ScienCell and PromoCell) were maintained in Endothelial Cell Medium (ECM, ScienCell) at 37 °C in a humidified air atmosphere with 5% CO2. All cells were used between passages 2 and 5. GL26 mouse glioma cells were kindly provided by Chunsheng Li (University of Pennsylvania) and cultured in Dulbecco's modified Eagle's medium (DMEM) medium (Gibco) supplemented with 5% fetal bovine serum (FBS). Chicken DF-1 fibroblasts (ATCC) were maintained in DMEM medium (ATCC, 30-2002) containing 5% FBS at 39 °C in a humidified air atmosphere with 5% CO2. T4123 glioma stem cells were kindly provided by Jeremy Rich (Cleveland Clinic) and then cultured in serum-free Neurobasal-A medium (Gibco), supplemented with B-27 Supplement Minus Vitamin A (Gibco), GlutaMax (Gibco), sodium pyruvate (Gibco), fibroblastic growth factor (5 ng/ml, R&D Systems), and EGF (20 ng/ml, R&D Systems). All cancer cell lines were checked and showed no mycoplasma contamination.
7
Hypoxia Stimulation of Multiple Myeloma Cells
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Human MM cell lines (RPMI, H929, and MM1.S) and Human stromal cells (HS5) were obtained from American Type Culture Collection (ATCC, LGC standards, Milan, Italy). All MM cells were routinely maintained in RPMI-1640 medium (Euroclone s.p.a., Milan, Italy); HS5 were maintained in DMEM medium (ATCC) and supplemented with 10% fetal bovine serum (FBS) and 1% Pen/strep (all from Euroclone s.p.a. Cells were maintained in a 37 °C humidified atmosphere of 5% CO2. To induce hypoxic stimulation, MM cell lines were seeded at 500,000 cells/ml and incubated in a “Hypoxic Chamber”containing 1% O2 gas mixture for 24 h. After hypoxia stimulation, cells were immediately kept on ice and processed.
8
RT4-D6P2T Cell Culturing and Drug Treatment
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RT4-D6P2T cell line was provided by ATCC (CRL-2768, batch 3993689, mycoplasma-free). Cells were thawed and cultured at Neurofit facilities (Illkirch, France) in DMEM medium (ATCC) containing 10% Foetal Bovine Serum (FBS; ATCC) and 1% antibiotic-antimicotic mixture (Gibco), and maintained in a humidified incubator at 37°C in 5% CO2-95% air atmosphere. Two days later, cells were trypsinised and transferred to 12-well plates (Nunc) at 30,000 cells per well. After 48 h, culture medium was replaced by DMEM containing drugs without FBS. After 8 h of treatment, cells were washed and harvested in 100 μL Phosphate Buffer Saline (PBS), then centrifuged 10 min at 13,000 rpm (4°C). The supernatant was discarded and the pellet was stored at −80°C until use. Three experiments with two independent cultures of RT4 were performed and each condition was done in triplicate.
9
Culturing and Authenticating Prostate Cell Lines
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22Rv1, VCaP, LNCaP, and 293T cells were obtained from ATCC. LNCaP95 cells were provided by Dr. Alan Meeker at Johns Hopkins University. VCaP and 293T cells were cultured in DMEM medium (ATCC, Catalog No. 30–2002) with 10% fetal bovine serum (Gibco, Catalog No.10437–028). LNCaP95 cells were cultured in phenol-red free RPMI–1640 medium (Gibco, Catalog No. 11835–030) with 10% charcoal-stripped fetal bovine serum (Atlanta, Catalog No. S11650). 22Rv1 and LNCaP cells were grown in RPMI-1640 medium (Hyclone, Catalog No. SH30027.02) with 10% fetal bovine serum. Cells used in all experiments were within 3 months of resuscitation of frozen cell stocks established within 3 passages after receipt of the cells. Cell authentication was performed at the Genetica DNA Laboratories, and cells were evaluated monthly for mycoplasma contamination.
10
Culturing Mouse Cancer Cell Lines
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Mouse CT26.WT colon, 4T1 breast and LLC-1 Lewis lung carcinoma cancer cell lines were purchased from an American Type Culture Collection (ATCC) local distributor, Sartorius (Beit Haemek, Israel). CT26 and 4T1 cell lines were grown in RPMI-based media (ATCC) supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin (Sartorius). LLC-1 cells were grown in DMEM medium (ATCC) supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin (Sartorius).
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