Human breast cancer cell line MCF-7 (tamoxifen-sensitive), T47D (tamoxifen-sensitive), BT-549, SK-BR-3, MDA-MB-231 and MCF-10A were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The identities of cell lines were confirmed by using DNA profiling (short tandem repeat, STR). MCF-7/TAM-1 and MCF-7/TAM-2 (tamoxifen-resistant) cell lines were established from MCF-7 cells after the following continuous exposure to 1 μM of TAM (Sigma, St. Louis, MO, USA) for more than 1 year. MCF-10A were cultured in DMEM/F12 (1:1) supplemented with 5% heat-inactivated equine serum, l0 μg/ml insulin 20 ng/ml EGF, 100 ng/ml cholera toxin, 0.5 μg/ml hydrocortisone and 2 mmol/l l -glutamine. BT-549 cells were cultured as previously described.42 (link) All other cell lines were maintained in DMEM medium supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) at 37 °C in a 5% CO2 humidified incubator. Cells were grown in monolayer and passaged routinely 2–3 times a week. Dimethyl sulfoxide (DMSO) and EST was purchased from Sigma.
Mcf 7
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Italy, Australia, China, India, Switzerland, Sao Tome and Principe
MCF-7 is a cell line derived from a breast adenocarcinoma. It is commonly used in laboratory research, particularly in the study of breast cancer and the effects of various compounds on breast cancer cell growth and behavior.
Automatically generated - may contain errors
Lab products found in correlation
Mcf 7, by American Type Culture Collection (150 mentions)
Mda mb 231, by Merck Group (74 mentions)
Mda mb 231, by American Type Culture Collection (71 mentions)
Fetal bovine serum (fbs), by Merck Group (47 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (32 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (31 mentions)
Rpmi 1640 medium, by Merck Group (29 mentions)
Penicillin streptomycin, by Merck Group (28 mentions)
Streptomycin, by Merck Group (26 mentions)
Penicillin, by Merck Group (25 mentions)
Mcf 10a, by American Type Culture Collection (23 mentions)
Rpmi 1640, by Merck Group (23 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (19 mentions)
Skbr3, by American Type Culture Collection (18 mentions)
Sk br 3, by Merck Group (18 mentions)
Bt474, by Merck Group (15 mentions)
L glutamine, by Merck Group (15 mentions)
Streptomycin, by Thermo Fisher Scientific (15 mentions)
Penicillin, by Thermo Fisher Scientific (15 mentions)
Mda mb 468, by Merck Group (15 mentions)
266 protocols using mcf 7
1
Establishing Tamoxifen-Resistant Breast Cancer Cell Lines
2
Establishment of Taxol-resistant Breast Cancer Cell Lines
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Human breast cancer cell lines MCF-7 and MDA-MB-231 (231) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The identities of cell lines were confirmed by using DNA profiling (short tandem repeat). Taxol-resistant cell lines MCF-7/Tax1, MCF-7/Tax2, 231/Tax1 and 231/Tax2 were established from MCF-7 and 231 cells after following continuous exposure to 2 and 1 nM of Taxol (Sigma, St. Louis, MO, USA) for >12 months. All cell lines were maintained in DMEM medium supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) at 37 °C in a 5% CO2 humidified incubator. Cells were grown in monolayer and passaged routinely 2–3 times a week. Dimethyl sulfoxide (DMSO) was purchased from Sigma.
3
Establishing Tamoxifen-Resistant Breast Cancer Cell Line
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The human breast cancer cell line MCF-7 was purchased from the Korean Cell Line Bank (Seoul, South Korea). MCF-7 cells are a well-characterized ER-positive control cell line. MCF-7 cells were seeded at a density of 2 × 105/cm2 and cultured in phenol-red-free RPMI 1640 medium containing 10% fetal bovine serum (FBS) and antibiotics. According to a methodology reported elsewhere [9 (link)], we established an MCF-7-derived tamoxifen-resistant cell line (TamR) via long-term culture of MCF-7 cells in the presence of 4-hydroxytamoxifen (Sigma-Aldrich, St. Louis, MO, USA). Briefly, MCF-7 cell monolayers were washed with phosphate-buffered saline (PBS) and transferred to phenol-red-free RPMI 1640 medium containing 10% charcoal-stripped, steroid-depleted FBS (Sigma-Aldrich), antibiotics, and 4-hydroxytamoxifen (10−7 M in ethanol). The cells were exposed to this treatment for 1 week, during which the medium was replaced twice a week. To generate drug-resistant cell lines, the cells were cultured in the presence of gradually increasing concentrations of 4-hydroxytamoxifen from 0.05 to 3 μM over a period of 8 months.
4
Indirect co-culture of MCF-7 cells and macrophages
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The ER+ and PR+ human breast cancer cell line MCF‐7 was purchased from DSMZ and maintained in RPMI‐1640 medium (PAA Laboratories Inc., Cölbe, Germany) supplemented with 10% fetal calf serum (FCS, Invitrogen, Karlsruhe, Germany). Human macrophages were derived from mononuclear peripheral blood cells and were collected with approval of the local ethics committee in Göttingen.
Briefly, monocytes were obtained from buffy coats of healthy donors through double‐density‐gradient isolation. Macrophages were differentiated by culturing monocytes in fluorinated ethylene propylene‐coated cell culture bags (CellGenix, Freiburg, Germany) in the presence of M‐CSF (Immunotools, Friesoythe, Germany) for 7 days. Differentiation into mature macrophages was assessed by flow cytometry verification of CD11b, CD11c, CD14 and CD45 (Beckman Coulter, Krefeld, Germany) expression and negativity for CD209 (BioLegend, Fell, Germany). For indirect co‐culture experiments, macrophages were seeded in hanging cell culture inserts (0.4 μm pore size, PET; Millipore, Billerica, MA, USA) and co‐cultured with MCF‐7 cells at a ratio of 2:1 under normoxia for 24 h. All co‐culture experiments were performed with at least 3 biological replicates, i.e. with different passages of MCF‐7 cells and macrophages derived from different donors.
Briefly, monocytes were obtained from buffy coats of healthy donors through double‐density‐gradient isolation. Macrophages were differentiated by culturing monocytes in fluorinated ethylene propylene‐coated cell culture bags (CellGenix, Freiburg, Germany) in the presence of M‐CSF (Immunotools, Friesoythe, Germany) for 7 days. Differentiation into mature macrophages was assessed by flow cytometry verification of CD11b, CD11c, CD14 and CD45 (Beckman Coulter, Krefeld, Germany) expression and negativity for CD209 (BioLegend, Fell, Germany). For indirect co‐culture experiments, macrophages were seeded in hanging cell culture inserts (0.4 μm pore size, PET; Millipore, Billerica, MA, USA) and co‐cultured with MCF‐7 cells at a ratio of 2:1 under normoxia for 24 h. All co‐culture experiments were performed with at least 3 biological replicates, i.e. with different passages of MCF‐7 cells and macrophages derived from different donors.
5
MCF-7 Cell Culture and Tamoxifen Induction
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The human BC cell line MCF-7 was bought from ATCC. MCF-7 cells were all maintained in Dulbecco's modified essential medium (DMEM), supplemented with 10% fetal bovine serum (FBS, Pricella, Wuhan, China), in a 5% CO 2 incubator. MCF-7-TAM was induced by 20 μM 4-hydroxytamoxifen (4-OHT, Calbiochem, Merck Millipore Billerica, MA, USA) incubation of MCF-7 for one month.
6
Establishing Tamoxifen-Resistant Breast Cancer Cells
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The tamoxifen-sensitive human breast cancer cell line MCF7 was obtained from the American Type Culture Collection (ATCC, HTB-22). Tamoxifen-resistant cells (MCF7/TAM) were established by culturing MCF7 cells in medium with 1 μM tamoxifen citrate salt (Sigma-Aldrich, T9262-1G) over 6 months, as previously described [30 (link), 31 (link)]. All of the cells were cultured in RPMI 1640 medium with 10% fetal bovine serum (FBS) at 37 °C in the presence of 5% CO2.
7
Breast Cancer Cell Lines Analysis
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The cell lines used in the analysis include MCF7, ZR-75-1, MCF7 Tam1, and ZR-75-1 Tam1, purchased from American Type Culture Collection (Rockville, MD, United States). Whereas ZR-75-1 and MCF7 were respectively cultured in 10% FBS-supplemented RPMI-1640 and EMEM, culture media for MCF7 was additionally contained and 10 μg/mL human insulin (Sigma. St. Louis, MO, United States). The tamoxifen-resistant cell lines, MCF-7 Tam1 and ZR-75-1 Tam1, used the same culture media as the parental cell lines, additionally containing 1 µM 4-hydroxytamoxifen (Sigma. St. Louis, MO, United States).
8
Establishment of Cisplatin-Resistant Breast Cancer Cell Lines
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MCF-7 and MDA-MB-231 cells were purchased from ATCC. In brief, the parental BC cells (defined MCF-7/Par and MDA-MB-231/Par) were continuously exposed to an ascending series of DDP (0.1~10 μM, Sigma-Aldrich, MO, USA) to induce DDP-resistant cells (MCF-7/DDP and MDA-MB-231/DDP). All cells were maintained in 10% FBS-supplemented DMEM containing 1% penicillin-streptomycin for further use.
9
Breast Cancer Cell Lines and Tamoxifen Resistance
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Human breast cancer cell lines MCF-7 (HTB-22, ATCC), T-47D (HTB-133, ATCC), ZR-75-1 (CRL-1500, ATCC) and BT-474 (HTB-20, ATCC) were obtained from the American Type Culture Collection. The cells were grown in DMEM with L-Glutamine (MCF-7 and BT-474, PAN Biotech, Aidenbach, Germany) or RPMI-1640 with L-Glutamine (ZR-75-1 and T-47D, PAN Biotech) supplemented with 10 % FCS (Gibco, Life Technologies, Carlsbad, CA) and 1 % penicillin/streptomycin (Gibco). Culture media for T-47D, MCF-7 and BT-474 additionally contained 0,1 % bovine insulin (Sigma. St. Louis, MO). The tamoxifen-resistant cell lines (MCF-7 Tam1, T-47D Tam1 & Tam2, ZR-75-1 Tam1 & Tam2, BT-474 Tam1 & Tam2) were derived from the parental cell lines by continuous exposure to 4-OH-tamoxifen (Sigma, 1 μM in ethanol) for 8–12 months. Culture media was replaced every 2–3 days. All cells were incubated at 37 °C with 5 % CO2 and passaged when ca 80 % confluent. The approximate doubling times of the cells were as follows: parental MCF-7, T-47D, ZR-75-1 and BT-474: 1–3 days. Resistant MCF-7 Tam1, T-47D Tam1 and Tam2: 1–2 weeks, ZR-75-1 Tam1 and Tam2: > 1 week, BT-474 Tam1 and Tam2: 2 weeks. The cells were free of mycoplasma and verified for their authenticity (Technology Centre, Institute for Molecular Medicine Finland, Helsinki, Finland).
10
Generating Tamoxifen-Resistant Breast Cancer Cells
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The breast cancer cell line MCF-7 was purchased from the American Type Culture Collection (Manassas, VA, U.S.A.) and routinely maintained in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum. Tamoxifen-resistant cells (MCF-7/TAM) were established by continuously culturing MCF-7 cells in the presence of 4 μM 4-hydroxy-Tamoxifen (Sigma–Aldrich, Missouri, U.S.A.) for 6 months. All cells were grown at 37°C in an atmosphere containing 5% CO2.
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