The cancer cell lines SUM-159 (breast cancer) and HT-29 (colorectal cancer) were plated in 96-well plates to assess the effects of duloxetine on OXA- and PTX-induced cell death using the MTT assay. SUM-159 cells were incubated in high glucose DMEM (Hyclone, United States) and HT-29 cells were incubated in RMPI-1640 medium (Gibco, United States) each containing 1% penicillin/streptomycin (Gibco, United States) and 10% FBS (Gibco, United States). Varying concentrations of duloxetine, 300 nM PTX, or 30 μM OXA were added to the wells. For the MTT assay, we added 10 μL of MTT (0.5 mg/mL) to each well. After 4 h the supernatant was removed and the formazan crystals were dissolved in 110 μL of DMSO. Absorbance was measured at 490 nm using a Multimode Plate Reader (Tecan, Switzerland).
Rmpi 1640 medium
Manufactured by Thermo Fisher Scientific
Sourced in United States, China, France
RPMI 1640 medium is a widely used cell culture medium formulation designed to support the growth of a variety of cell types, including human and animal cells. It provides a balanced salt solution, amino acids, vitamins, and other nutrients required for cell proliferation and maintenance in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (56 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (14 mentions)
Streptomycin, by Thermo Fisher Scientific (12 mentions)
Dmem medium, by Thermo Fisher Scientific (9 mentions)
Penicillin, by Thermo Fisher Scientific (9 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (7 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (7 mentions)
Lipopolysaccharides (lps), by Merck Group (5 mentions)
Hep 2, by Cell Resource Center (4 mentions)
Tu212, by Cell Resource Center (4 mentions)
Thp 1, by American Type Culture Collection (3 mentions)
Mem medium, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Sartorius (3 mentions)
Normal human sera, by Merck Group (2 mentions)
Skov3, by American Type Culture Collection (2 mentions)
Thp 1, by Thermo Fisher Scientific (2 mentions)
Hek293t, by American Type Culture Collection (2 mentions)
Mcf 7, by American Type Culture Collection (2 mentions)
Tib 202, by American Type Culture Collection (2 mentions)
108 protocols using rmpi 1640 medium
1
Duloxetine's Effects on Chemotherapy-Induced Cell Death
2
Polymer-Peptide Conjugates for Targeted Cancer Therapy
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We obtained the methoxy poly (ethylene glycol) 3000-poly (lactic acid) 34000 (MPEG-PLA) and carboxyl-poly (ethylene glycol) 3400-poly (lactic acid)34000 (HOOC-PEG-PLA) from LACTEL Absorbable Polymers (USA). 1, 10-Dioctadecyl-3,3, 3-tetramethylindo-tricarbocyanineiodide (DiR) was purchased from Biotium (Invitrogen, USA). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetra-zoplium bromide (MTT) and coumarin-6 were purchased from Beyotime (Haimen, China). RMPI 1640 medium, fetal bovine serum (FBS) and trypsin-EDTA solutions were obtained from Gibco (CA, USA). All peptides were synthesized by Shanghai Mocell Biotech Co., Ltd (China). Sequences of each peptide used here was CREKA-NH2 for C peptide, CKDEPQRRSARLSAKPAPPKPEPKPKKAPAKK-NH2 for F3 peptide, and CREKA-CN2-CKDEPQRRSARLSAKPAPPKPEPKPKKAPAKK-NH2 for CF peptide. All other reagents were of analytical grade and were from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China).
The MDA-MB-231 cell line was obtained from the Chinese Academy of Sciences Cell Bank. Female nude Balb/c mice (18–20 g) were purchased from Shanghai Sino-British Sippr/BK Lab Animal Ltd. (Shanghai, China) and raised under the standard condition. All the animal experiments were conducted in accordance with the guidelines of the Second Military Medical University.
The MDA-MB-231 cell line was obtained from the Chinese Academy of Sciences Cell Bank. Female nude Balb/c mice (18–20 g) were purchased from Shanghai Sino-British Sippr/BK Lab Animal Ltd. (Shanghai, China) and raised under the standard condition. All the animal experiments were conducted in accordance with the guidelines of the Second Military Medical University.
3
ConA-Induced PBMC Proliferation Assay
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Fresh peripheral blood mononuclear cells (PBMCs) were obtained from whole blood by Ficoll-Hypaque density gradient. PBMCs were placed in a 96-well-plate at 3.0 × 105 cells per well in RMPI 1640 medium (Gibco, Gaithersburg, MD) with 10% normal human sera (Sigma, St. Louis, MO); 5 μg of concanavalin A (Sigma-Aldrich) was added, and cells were incubated at 37°C and 5% CO2 for 5 days. EdU Click-iT Alexa Fluor 488 (Invitrogen) was added for 6 h, and cells were acquired by flow cytometry using the FACSCanto II cytometer (Becton Dickinson); the lymphoproliferation was analyzed using the FacsDiva software (Becton Dickinson) determined by the difference in fluorescence intensity means (MiFID) of the control and ConA wells.
4
Authentication and Culturing of HGSOC Cell Lines
5
Cell Culture of Human Cancer Lines
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Two human ovarian cancer cell lines (A2780, SKOV3), a human cervical adenocarcinoma cell line (Hela) and a human cervical squamous cell carcinoma cell line (Siha) were obtained from the American Type Culture Collection (ATCC, USA).29–32 (link) A2780 and SKOV3 cell lines were maintained in RMPI 1640 medium (Gibco, USA) supplemented with 10% FBS (Biological Industries, Israel). Hela cell lines were cultured in the DMEM medium (Gibco, USA) enriched with 10% FBS. And Siha cell lines were maintained in MEM medium (Gibco, USA) supplemented with 10% FBS. All the cell lines were cultured under the condition of 37°C and 5% CO2.
6
Propagation of Neuroblastoma Cell Lines
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Mouse neuroblastoma (NA) cells were maintained in RMPI 1640 medium (Gibco, USA) which contains 10% fetal calf serum (FBS). CVS-11 and HEP-Flury were propagated in NA cells. Newcastle disease virus (NDV) vaccine strain LaSota was purchased from Winsun Pharm Company (Guangzhou, China). Female BALB/c mice were purchased from Center for Laboratory Animal Science, Southern Medical University (Guangzhou, China) and kept at the Laboratory Animal Center of South China Agricultural University.
7
Differentiation of THP-1 Cells into Foam Cells
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The human monocytic cell line THP-1 and human embryonic kidney (HEK)293T cells were purchased from the American Type Culture Collection (ATCC). THP-1 cells were cultured in RMPI-1640 medium (Gibco, Carlsbad, CA, USA), and HEK-293T cells were cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco). Both media were supplied with 10% fetal bovine serum (Gibco), 100 μg/mL streptomycin, and 100 IU/mL penicillin (Thermo Fisher Scientific, Rockford, IL, USA). Cells were incubated in a humidified incubator at 37 °C with 5% CO2. To drive monocyte differentiation into macrophages and ultimately establish a foam cell model, THP-1 cells were first seeded at 1 × 106 per mL with 100 nM propylene glycol monomethyl ether acetate (PMA, Sigma-Aldrich, St. Louis, MO, USA) for 12 hours to differentiate them into adherent macrophages18 (link)50 (link). The THP-1 macrophages were then stimulated with 50 μg/ml oxLDL (Luwen, Shanghai, China) for 12 hours to establish the foam cell model51 (link).
8
Culturing Colorectal Cancer Cell Lines
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Human CRC HCT8S and HCT8R were purchased from Shanghai Meixuan Corporation (Shanghai, China). HCT8S cell lines were cultured in Dulbecco’s Modified Eagle’s medium (GIBCO, Carlsbad, CA, U.S.A.) supplemented with 10% fetal bovine serum (GIBCO, Carlsbad, CA, U.S.A.). HCT8R cell lines were cultured in RMPI 1640 medium (GIBCO, Carlsbad, CA, U.S.A.) with 10% fetal bovine serum and 15 μg/ml 5-Fu (Sigma-Aldrich, Northbrook, IL, U.S.A.) according to the manufacturer’s protocols. Both kinds of cells were maintained in an atmosphere containing 5% CO2 at 37°C according to a previous report [22 (link)].
9
Cell Culture and Compound Preparation
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Non-small-cell lung cancer (A549), oral epidermoid carcinoma (KB), cervical cancer (Hela) and breast cancer (MCF-7) cells were purchased from the American Type Culture Collection (ATCC, America). Human umbilical vein endothelial cell (HUVEC) and human normal hepatocytes (LO2) cells were provided by Guangzhou Medical University (Guangzhou, China). A549 and KB cells were cultured in RMPI-1640 medium (Gibco, America). Hela, MCF-7 and LO2 cells were cultured in DMEM medium (Gibco, America). HUVEC cells were cultured in Ham’s F-12K medium (Gibco, America). All the mediums were supplemented with 10% fetal bovine serum (FBS) (BI, State of Israel), 100 U/mL penicillin and 100 μg/mL streptomycin (Gibco, America), and all the cells were incubated in a sterile incubator at 37°C in an environment of 5% CO2. The tested compounds were dissolved in dimethyl sulfoxide (DMSO) at a concentration of 10 mM, and then diluted into different concentration of solutions by culture medium. The final concentration of DMSO was <0.01% (v/v) in all drug treatment groups to avoid the toxicity in the biological system. Untreated cells were used as a blank control and 5-fluorouracil treated cells were used as a positive control.
10
Macrophage Generation from Human Monocytes
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Peripheral blood mononuclear cells (PBMCs) were isolated from patients or buffy-coats from healthy donors over a ficoll gradient (Lymphocytes Isolation Solution, Rafer). Monocytes were purified by magnetic cell sorting using anti-CD14 tagged microbeads (Miltenyi Biotech, Bergisch Gladbach, Germany). For in vitro generation of macrophages, monocytes were cultured at 0.5 × 106/mL for 7 days containing GM-CSF (10 ng/mL or 1,000 U/mL, Immunotools) or M-CSF (10 ng/mL, Immunotools) to generate MP-GM or MP-M macrophages, respectively. Cytokines were added every two days. All cells, including melanoma cell lines BLM and A375 [21 (link)] were cultured in RMPI-1640 medium (Gibco, Waltham, MA, USA) supplemented with 10% fetal calf serum (FCS, Sigma, Burlington, MA, USA).
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