ZR75.1 (ATCC-CRL1500), luminal B subtype human breast cancer cell line was used in our experiments. Cells were grown in 10% foetal bovine serum (FBS, Biosera), glutamine (2 mM) and gentamycin-contained RPMI-1640 media (Biosera—Nuaille, France) at standard cell culture conditions. Different treatments were applied in 96-well plates, tissue culture flasks, and 3D bioprinted tissue-mimetic scaffolds. Before the treatments, the bioprinted scaffolds were maintained for 7 days, while the cells in 2D cell cultures were incubated for 24 h. After media refreshment, the 72-h treatments were carried out in 96-well plates, tissue culture flasks, and on scaffolds (minimum 6 parallels maintained in every well of 6-well plates) for sensitivity tests. mTORC1 inhibitor rapamycin (Rapa; 50 ng/mL; Focus Biomolecules, Plymouth Meeting, PA, United States), the natural anthracycline antibiotic and chemotherapeutic agent doxorubicin (Doxo; 50 ng/mL; TEVA, Debrecen, Hungary) and their combinations were applied regarding our previous mTOR inhibitor combination sensitivity studies (102 (link)).
Rpmi 1640 media
Manufactured by Biosera
Sourced in France
RPMI-1640 media is a cell culture medium commonly used for the in vitro cultivation of a variety of cell types, including human and animal cells. It is a balanced salt solution that provides the necessary nutrients and growth factors to support the growth and maintenance of these cells.
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Lab products found in correlation
5 protocols using rpmi 1640 media
1
Breast Cancer Cell Line Treatment Evaluation
2
CXCR4 Expression on Cell Lines
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Selected cell lines were purchased from Iran Cell Collection Bank (Pasteur Institute). These cell lines were cultured in RPMI 1640 media (Bio sera) with 10% FBS (GIBCO), 100 IU/mL penicillin, and 100 µg/mL streptomycin (GIBCO). To characterize the expression of CXCR4, the cell lines were stained with anti-CXCR4 antibody and analyzed by flow cytometry. Then, after optimization of the incubation time for coculture, cell lines were incubated (1×105 cell/mL) with the PMPs (10, 20, 50 125, 250, 500, 1000 μ g/mL) for 1 hour in 5% CO2 Incubator (37˚C). All tests were repeated 3 times for all the cell lines before and after treatment with PMPs.
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Selected cell lines were purchased from Iran Cell Collection Bank (Pasteur Institute). These cell lines were cultured in RPMI 1640 media (Bio sera) with 10% FBS (GIBCO), 100 IU/mL penicillin, and 100 µg/mL streptomycin (GIBCO). To characterize the expression of CXCR4, the cell lines were stained with anti-CXCR4 antibody and analyzed by flow cytometry. Then, after optimization of the incubation time for coculture, cell lines were incubated (1×105 cell/mL) with the PMPs (10, 20, 50 125, 250, 500, 1000 μ g/mL) for 1 hour in 5% CO2 Incubator (37˚C). All tests were repeated 3 times for all the cell lines before and after treatment with PMPs.
3
Biological Effects of Cu(II) Complexes on Cancer Cells
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To test the biological effects of the Cu(II) complexes and the ligands on human cancer cells, DU-145 prostate, A549 lung, and MCF-7 breast adenocarcinoma cells, as well as the multidrug-resistant MCF-7 KCR breast cancer cells were utilized. All the cell lines were maintained in RPMI-1640 media (Biosera, Nuaille, France) complemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 0.01% streptomycin and 0.005% penicillin, (Biosera, Nuaille, France) and were cultured under standard conditions in a 37 °C incubator at 5% CO2 and 95% humidity. The cell lines were originally obtained from ATCC.
4
Culturing K562 Cells for NK Cytotoxicity
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K562 cell line (a human erythroleukemia cell line [acute myeloid leukemia-M6 {AML}]) was purchased from Pasteur Institute of Iran. Before NK cells cytotoxicity experiments, K562 cells were cultured in complete RPMI-1640 media (Biosera, Nuaille, France) supplemented with 10% fetal bovine serum (Biosera, Nuaille, France), 2 mM L-glutamine, 100 IU/ml penicillin, and 100 μg/ml streptomycin.
5
Bystander Effect in Lung Cancer Cells
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Human lung carcinoma (QU-DB) cell line was obtained from the Pasteur Institute, Tehran, Iran. It was treated as target and bystander cells. The cells were grown in RPMI-1640 media (Biosera, England) supplemented with 10% fetal bovine serum, 100 μg/ml streptomycin, and 100 U/ml penicillin. The cells were incubated at 37°C in a humidified atmosphere of 95% air and 5% CO 2 . Two days before to irradiation, QU-DB cells were trypsinized and cultured in (2.5 × 10 5 ) 12 cm 2 flasks. Two main groups were defined: Target and bystander groups. Two hours before irradiation, the culture media of target flasks were replaced with fresh medium. Irradiation was performed with a 60 Co teletherapy unit (Theratron, phoenix model, average dose-rate of 60/79 cGy/min) at doses of 2, 4, 6, and 8 Gy. The radiation field size was 15 cm × 15 cm and source to medium distance was 80 cm. Following irradiation, target flasks were returned to the incubator. After 1 h, irradiated culture medium extracted from irradiated flasks was filtered through 0.22 μm acetate cellulose filter (Orange Scientific, Belgium) to remove any dead cells. Then, they were transferred to specified bystander flasks. Following to medium transfer, all groups including irradiated, sham-irradiated and bystander flasks were incubated for 24 h.
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