MCF7 breast cancer cell line was obtained from ATCC. MCF7 cells were grown in DMEM medium, with 10% fetal bovine serum (FBS), penicillin (100 U/mL) and streptomycin (100 µg/mL). Cells were plated 24 h before treatment with different drugs at the indicated concentrations.27 (link) Recombinant human heregulin (HRG) and IGF1 were obtained from Sigma Chemical Co (St. Louis, MO).27 (link) PRCP inhibitor (PRCP-7414); catalog number 504044 was from Calbiochem. OSI906 and lapatinib are obtained from Selleckchem.
Mcf 7 breast cancer cell line
Manufactured by American Type Culture Collection
Sourced in United States
The MCF-7 breast cancer cell line is a well-established in vitro model derived from a human breast adenocarcinoma. It is a commonly used cell line in cancer research and drug discovery studies.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (11 mentions)
Streptomycin, by Thermo Fisher Scientific (6 mentions)
Penicillin, by Thermo Fisher Scientific (6 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Lonza (2 mentions)
Dmem f12, by Thermo Fisher Scientific (2 mentions)
Htert bj1, by Takara Bio (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Trypsin edta solution, by Thermo Fisher Scientific (2 mentions)
Igf 1, by Merck Group (1 mentions)
Lapatinib, by Selleck Chemicals (1 mentions)
Osi 906, by Selleck Chemicals (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
Streptavidin hrp, by R&D Systems (1 mentions)
Vybrant cell labeling solution, by Thermo Fisher Scientific (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Annexin 5 alexa 488, by Thermo Fisher Scientific (1 mentions)
Propidium iodide, by Santa Cruz Biotechnology (1 mentions)
41 protocols using mcf 7 breast cancer cell line
1
Breast Cancer Cell Line MCF7 Treatment Protocols
2
Multimodal Profiling of Cell Lines
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The MCF7 breast cancer cell line, HEK-293 and HEK-293T embryonic kidney cell lines, and monocytic lymphoma cell line U937 were purchased from ATCC® (Manassas, Virginia, USA). The mouse anti-CD47 mAb B6H12 was either produced and purified from hybridoma HB9771 purchased from ATCC®, or purchased from Santa-Cruz Biotechnology (Dallas, TX, USA). The biotinylated mouse anti-CD47 mAb (clone B6H12.2) was purchased from BioLegend (San Diego, CA, USA). The monoclonal sheep anti-SIRPa antibodies and streptavidin-HRP were purchased from R&D Systems (Minneapolis, MN, USA). The monoclonal anti-M13-HRP antibody was from Thermo (Waltham, MA, USA). All the secondary antibodies were purchased from Santa-Cruz Biotechnology. The recombinant human SIRPa-Fc was from R&D Systems. The human CD47 with His tag was a kind gift from P.M. Chumakov. The streptavidin was purchased from MyBioSource (San Diego, CA, USA). The fluorescent lipophilic dyes DiO and DiL were from Biotium (San Francisco Bay Area, CA, USA). The Annexin V-Alexa 488 was from Invitrogen (Waltham, MA, USA), while the propidium iodide was from Santa-Cruz Biotechnology. The Hoechst 33342 was purchased from Sigma (Burlington, MA, USA). The DiL and DiO dyes were from Vybrant Cell-Labeling Solutions, Invitrogen.
3
MCF-7 Breast Cancer Cell Culture Protocol
4
Cell Culture Maintenance Protocol
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The MCF7 breast cancer cell line, HCT 116 colorectal carcinoma cell line, A-704 kidney adenocarcinoma cell line, and WI-26 VA4 lung epithelial-like cells were purchased from the ATCC. The MCF7 cells were maintained in MEM (Gibco, Paisley, UK) supplemented with 10% fetal bovine serum (FBS, Gibco, UK), human recombinant insulin (0.01 mg/mL), penicillin (100 UI mL−1), streptomycin (100 µg mL−1), and GlutaMax (2 mM, Gibco, UK). The HCT 116 cells were maintained in McCoy’s 5A (Gibco, UK) supplemented with 10% fetal bovine serum (FBS, Gibco, UK), penicillin (100 UI mL−1), streptomycin (100 µg mL−1), and GlutaMax (1.5 mM, Gibco, UK). A-704 and WI-26 VA4 cells were maintained in MEM (Gibco, UK) supplemented with 10% fetal bovine serum (FBS, Gibco, UK), penicillin (100 UI mL−1), streptomycin (100 µg mL−1), and GlutaMax (1.87 mM, Gibco, UK). All cell lines’ cultivation was performed under a humidified atmosphere of 95% air/5% CO2 at 37 °C. Subconfluent monolayers, in the log growth phase, were harvested by a brief treatment with TrypLE Express solution (Gibco, UK) in phosphate-buffered saline (PBS, Capricorn Scientific, Ebsdorfergrund, Germany) and washed three times in serum-free PBS. The number of viable cells was determined by trypan blue exclusion.
5
Comprehensive Cell Line Profiling
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Five cell lines were selected to include a range of TF expression and not on the basis of tissue of origin. BxPC-3 pancreatic cancer and 786-O renal carcinoma cell lines (ATCC, Teddington, United Kingdom) were cultured in RPMI-1640 medium, MCF-7 breast cancer cell line and HepG2 hepatocellular carcinoma cell lines (ATCC) were cultured in EMEM, and MDA-MB-231 (ATCC) breast cancer cell lines were cultured in DMEM. All media were supplemented with 10% (v/v) heat-inactivated fetal calf serum (FCS) to ensure the lack of any functional enzymes. Human coronary artery endothelial cells (HCAECs), devoid of endogenous TF, were cultured in MV media containing 5% (v/v) FCS and growth supplements (PromoCell, Heidelberg, Germany). In some experiments, the cells were adapted to serum-free medium prior to use.
6
Stable Transduction of MCF7 Breast Cancer Cells
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The MCF7 breast cancer cell line was purchased from ATCC. MCF7 breast cancer cells were stably transduced with lentiviral vectors (i) pLENTI6.3-RBS-COMT-IRES-EmGFP-GWs expressing COMT (without tag) and green fluorescence protein (GFP) (MCF7-COMT), (ii) pLENTI6.3-EmGFP-GWs expressing GFP as a control (MCF7-GFP), (iii) pLENTI6.3-N-SBP-TEV-COMT-IRES-EmGFP-GWs expressing COMT N-terminally tagged with streptavidin binding peptide and GFP (MCF7-SBP-COMT) and (iv) pLENTI6.3-N-SBP-EmGFP-GWs expressing N-terminal SBP and GFP (MCF7-SBP) as described in Supplementary Methods. The parental MCF7 cells were cultured in Dulbecco's Modified Eagle´s Medium (DMEM; high glucose) with 10% fetal bovine serum (Merck, Germany) at 37 °C in 5% CO2 atmosphere; the stably transduced MCF7 cells were cultured under the same conditions except for adding blasticidin (10 μg/ ml; InvivoGen, USA) to the culture medium after the first passage.
7
MCF7 Breast Cancer Cell Line REAC-RGN Treatment
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The MCF7 breast cancer cell line was acquired from ATCC (Manassas, VA, USA). The cells were cultured in a medium composed of Dulbecco’s modified Eagle’s medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA), 10% fetal bovine serum (FBS, Thermo Fisher Scientific, USA), 200 mM L-glutamine (Thermo Fisher Scientific) and 200 U/mL penicillin−0.1 mg/mL streptomycin (Thermo Fisher Scientific, USA). The REAC-RGN treatment is a pre-programmed treatment. Cells were cultured in the absence or presence of REAC- RGN treatment for different time points.
A group of cells was exposed to REAC RGN treatment for 72 h and then put in culture for additional 4 days (a total of 7 days).
A second group of cells was exposed to REAC RGN treatment for 72 h and then put in culture for additional 7 days (a total of 10 days).
A third group of cells was exposed to REAC RGN treatment for 7 days and then put in culture for additional 7 days (a total of 14 days). For each group, the untreated controls were represented by cells cultured in the growing medium alone (Figure 1 ). The experiments were performed two times with three technical replicates for each sample.
A group of cells was exposed to REAC RGN treatment for 72 h and then put in culture for additional 4 days (a total of 7 days).
A second group of cells was exposed to REAC RGN treatment for 72 h and then put in culture for additional 7 days (a total of 10 days).
A third group of cells was exposed to REAC RGN treatment for 7 days and then put in culture for additional 7 days (a total of 14 days). For each group, the untreated controls were represented by cells cultured in the growing medium alone (
8
MCF-7 Breast Cancer Cell Protocol
9
Cell Culture Protocols for Cancer Research
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MCF7 breast cancer cell line was obtained from the ATCC. The cells were cultured using the same protocol reported by Azzi et al42 (link). Briefly, cells were seeded in flasks using EMEM medium (Lonza) supplemented with 10% (v/v) fetal bovine serum (FBS) (Lonza), 1 mM sodium pyruvate, 2 mM glutamine, non-essential amin o acids, 0.01 mg/mL human insulin (Sigma Aldrich). Mouse mesothelioma (AB22) cell line was obtained from Sigma Aldrich and they were cultured in RPMI medium (Lonza) supplemented with 25 mM Hepes, 10% (v/v) FBS and 2 mM glutamine. Human mesothelioma (ZL34) cell line was obtained from Sigma Aldrich. The cells were cultured in DMEM-Ham’s F12 (Lonza) containing 2.5 mM glutamine and supplemented with 15% (v/v) FBS using the same protocol described in reference35 (link). All media contained 100 U/mL penicillin and 100 U/ml streptomycin. All the cell lines were maintained in a humidified incubator at 37 °C, 5% CO2.
10
Culturing MCF7 and hTERT-BJ1 Cells
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MCF7 breast cancer cell lines were purchased from the ATCC. Human immortalized fibroblasts (hTERT-BJ1) were originally purchased from Clontech, Inc. Cells were cultured in Dulbecco's modified Eagle's medium (DMEM), supplemented with 10% FBS (fetal bovine serum), 2 mM GlutaMAX, and 1% Pen-Strep in a 37°C humidified atmosphere containing 5% CO2, unless other-wise noted. Gibco-brand cell culture media (DMEM/F12) was purchased from Life Technologies. Bedaquiline was purchased from AdooQ Bioscience.
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