Normal epithelial breast cell line MCF-10A and breast cancer cell line MCF-7 were purchased from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai Institute of Cell Biology, Shanghai, China). MCF-10A cells were maintained in a mixed medium as per Qin's method 9 (link). adriamycin-resistant MCF-7/ADR cells were screened from MCF-7 cells in the optimal growth state after being exposed to different concentrations of adriamycin (MedChemExpress, Shanghai, China). MCF-7 cells were cultured in DMEM F12 supplemented with 10% FBS. MCF-7/ADR cells were cultured in RPMI 1640 media supplemented with 10% FBS. All media were supplemented with 100 unit/mL penicillin, and 100 μg/mL streptomycin, and all cells were incubated at 37 °C in a 5% CO2 atmosphere. Specifically, to sustain the multidrug resistant phenotype, MCF-7/ADR cells were cultured in the medium supplemented with adriamycin (4 μg/ml). Before in vitro assay, all cells were cultured in a drug-free medium for 24 hrs.
Mcf 7
Manufactured by MedChemExpress
Sourced in China, United States
MCF-7 is a human breast adenocarcinoma cell line commonly used in research applications. It is a well-established model for studying breast cancer biology and evaluating the effects of various compounds on breast cancer cells.
Automatically generated - may contain errors
Lab products found in correlation
Tamoxifen, by MedChemExpress (2 mentions)
Mda mb 231, by MedChemExpress (2 mentions)
Palbociclib, by MedChemExpress (1 mentions)
Arachidonic acid, by Merck Group (1 mentions)
Cisplatin, by MedChemExpress (1 mentions)
Doxorubicin, by MedChemExpress (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Beyotime (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Bt549, by American Type Culture Collection (1 mentions)
Mda mb 468, by American Type Culture Collection (1 mentions)
Hs578t, by American Type Culture Collection (1 mentions)
Mda mb 436, by American Type Culture Collection (1 mentions)
Rpmi 1640 medium, by Sartorius (1 mentions)
Hek293t, by Sartorius (1 mentions)
Tamoxifen hy 13757a, by MedChemExpress (1 mentions)
Mcf 7, by Sartorius (1 mentions)
Dulbecco modified eagle medium (dmem), by Sartorius (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
5 protocols using mcf 7
1
Culturing Adriamycin-Resistant Breast Cancer Cells
2
Generating Palbociclib-Resistant MCF-7 Cells
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The MCF-7 was purchased in 2017 to 2018 from the Chinese Academy of Science Committee Type Culture Collection Cell Bank (Shanghai, China). MCF-7 cells were cultured in DMEM (Dulbecco's modified Eagle's medium) high glucose supplemented with 10% fetal bovine serum (FBS) and 100 units/ml of penicillin and 100 units /ml of streptomycin. The MCF-7 cells were maintained in a humidified atmosphere with 5% CO2 at 37 °C.
MCF-7 cells were initially cultured in medium containing 1 µM palbociclib (MedChem Express, Shanghai, China), and then cells were subcultured every 2-3 weeks in medium with increased concentrations of palbociclib (a 25% increase each time). Finally, the obtained cells grew exponentially in the presence of 30 µM palbociclib and were named as MCF-7pR.
MCF-7 cells were initially cultured in medium containing 1 µM palbociclib (MedChem Express, Shanghai, China), and then cells were subcultured every 2-3 weeks in medium with increased concentrations of palbociclib (a 25% increase each time). Finally, the obtained cells grew exponentially in the presence of 30 µM palbociclib and were named as MCF-7pR.
3
Establishing Drug-Resistant Breast Cancer Cell Lines
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The human breast cancer cells (BT-549, Hs578T, MDA-MB-231, MDA-MB-468, MDA-MB-436, MCF-7, and T47D) were obtained from the American Type Culture Collection (ATCC, USA). All cells were cultured in the recommended medium (Gibco, USA), supplemented with 10% fetal bovine serum (Gibco, USA) and 1% streptomycin/ penicillin (Beyotime, Shanghai, China) at 37 °C with 5% CO2. To establish cisplatin (DDP)-resistant MDA-MB-231 cell line (MDA-MB-231/DDP), doxorubicin-resistant BT549 cell line (BT-549/DOX) and tamoxifen-resistant MCF-7 cell line (MCF-7/TAM), MDA-MB-231, BT-549 and MCF-7 cells were exposed to repetitive and incremental concentrations of drugs over a period of 6 months. To maintain the resistance phenotype, MDA-MB-231/DDP, BT-549/DOX, and MCF-7R cells were, respectively, cultured in the presence of 2 μM cisplatin (Med-ChemExpress, NJ, USA), 2 μM doxorubicin (Med-ChemExpress, NJ, USA), and 1 μM tamoxifen (Med-ChemExpress, NJ, USA). Arachidonic acid was obtained from Sigma-Aldrich (St. Louis, MO, USA). Giripladib, an inhibitor of cytoplasm phospholipase A2, was purchased from USBiological Life Sciences (Swampscott, MA, USA).
4
Culturing Tamoxifen-Resistant Breast Cancer Cells
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MCF7, T47D and HEK293T cells were obtained from the American Type Culture Collection (ATCC). MCF7, Tamoxifen-resistant MCF7 and HEK293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; 01-052-1ACS, Biological Industries) supplemented with 10% fetal bovine serum (FBS; 04-001-1ACS, Biological Industries), and T47D cells were cultured in RPMI 1640 medium (01-100-1ACS, Biological Industries) supplemented with 10% FBS. Tamoxifen-resistant MCF7 cells were developed by culturing MCF7 cells in the presence of 2 μM Tamoxifen (HY-13757A, MedChemExpress) for >12 months. Tamoxifen-resistant MCF7 cells were then maintained in the presence of 1 μM Tamoxifen. All cells were cultured in a humidified incubator with 5% CO2 at 37°C. If estrogen (E2, E8875, Sigma) was added, cells were maintained in stripping medium (phenol red free) plus 5% charcoal-depleted FBS for 72 h before treatment.
5
Breast Cancer Cell Lines Treatment
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The MCF-10A (cat. no. CRL-10317, Manassas, Virginia, USA), MCF7 (cat. no. CL-0149, Manassas) and MDA-MB-231 (cat. no. CL-0150B, Manassas) cell lines were obtained from the American Type Culture Collection through commercial channels. MCF7 cells were cultured in DMEM (PM150210, Procell, Wuhan, China) containing 10% fetal bovine serum (FBS) (Procell), penicillin (50 U/mL, 10378016, Gibco, Waltham, MA, USA), and streptomycin (50 µg/mL, 10378016, Gibco) in 5% CO2. MDA-MB-231 cells were cultured in F15 medium (PM150110, Procell) containing 15% FBS, penicillin (50 U/mL), and streptomycin (50 µg/mL) in 5% CO2. MCF7 and MDA-MB-231 cells were cultured in sinensetin (HY-N0297, MedChemExpress, New Jersey, USA) at different concentrations (0, 30, 60, 120, 180, and 240 µM) for 24 h. Cells were treated with SKL2001 (SKL101085, MedChemExpress) for one hour before the exposure of sinensetin according to the previous study (32 (link)). The concentration of 40 µM SKL2001 was in accordance with the previous study (33 (link)).
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