Cell lines were maintained at 37 °C at 5% CO2 in RPMI 1640 or DMEM culture media (Sigma) supplemented with 10% fetal bovine serum (Sigma), 50 Uml−1 penicillin and 50 µgml−1 streptomycin (Life Technologies). MEL526, PC3 cell lines in RPMI 1640, and HT108, RPMI 8322, MEL2664 cell lines in DMEM maintained. Hypoxic conditions (1% O2) were achieved in a Ruskinn InvivO2 400 hypoxia chamber, by supplementing ambient air with balanced N2 and CO2.
Dmem culture media
Manufactured by Merck Group
Sourced in United States
DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture media developed by Merck Group. It is a commonly used basal medium that provides essential nutrients, vitamins, and other components required for the in vitro growth and maintenance of various cell lines.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
Streptomycin sulfate, by Merck Group (1 mentions)
Antibiotic antimycotic, by Merck Group (1 mentions)
Penicillin g, by Merck Group (1 mentions)
Amphotericin b, by Merck Group (1 mentions)
3 protocols using dmem culture media
1
Cell Line Maintenance and Hypoxic Culture
2
Interaction of HT1080 cells with Fe7C3@C MNPs
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Human fibrosarcoma cells HT1080 (kindly provided by Russian Collection of cell lines, St. Petersburg) were cultured in DMEM culture media (Sigma, USA) supplemented with 10 % fetal calf serum (HyClone, USA) and antibiotic–antimycotic (100 units/ml penicillin G, 100 mg/ml streptomycin sulfate and 0.25 mg/ml amphotericin B) (Sigma, USA). For microscopic experiments cells were plated onto cover slips at a concentration of 10,000 cells/cm2 and grown for 48 h to reach 50 % confluency before the addition of Fe7C3@C MNPs to a final concentration of 20 µg/ml. Kinetics of cell interaction with Fe7C3@C was studied by TEM on serial ultrathin sections, and also by optical microscopy using time lapse video recording of living cells.
3
Glioma Cell Apoptosis Regulation
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Human glioma cells LN-18 were purchased from American Type Culture Collection (ATCC). MTT kit was purchased from Shanghai Bioleaf Technology Co. (Shanghai, China). Apoptosis detection kits including caspase-3 activity assay kit and FITC-Annexin-V apoptosis detection kit for phosphatidylserine (PS) externalization were purchased from Beijing Dingguo Biotechnology Co. (Beijing, China). Anti-Tim4 and anti-actin antibodies were purchased from Beyotime Biotechnology (Shanghai, China). DMEM culture media and fetal bovine serum (FBS) were purchased from Sigma (St. Louis, MO, USA). Antisense miRNA targeting Tim4 and control miRNA were designed and synthesized by Fengshou Biotechnology Co. (Shanghai, China). Lipofectamine 2000 transfection reagent was purchased from Shanghai Bioleaf Technology Co. The collection of human tissues was approved by the Research Ethics Committee at Qilu Hospital and performed in strict accordance with international standards. All patients were required to sign the informed consent.
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