Human breast cancer cell lines (MDA-MB-231, MCF-7, ZR-75-1) were purchased from the ECACC culture collection (Sigma Aldrich, Wicklow, Ireland). Cell lines were routinely tested upon freezing and thawing for mycoplasma contamination using PCR. Cells were maintained at 37 °C, 5% CO2 humidified incubator in DMEM-high glucose (MDA-MB-231 & MCF-7) or RPMI (ZR-75-1) supplemented with 10% fetal bovine serum, 5% l-glutamine and 5% penicillin/streptomycin (all obtained from Sigma Aldrich). ZR-75-1 media was additionally supplemented with 5% sodium pyruvate (Sigma Aldrich). For 3-dimensional culture, the on-top method was used where cells were seeded into wells containing Growth-Factor Reduced Matrigel™ (Biosciences, Dublin, Ireland) then covered with DMEM supplemented with 2% Matrigel. For analysis of cell signaling, cells were washed with PBS before being covered with serum free medium for 4 h prior to EGF stimulation (Peprotech, London, UK).
Zr 75 1
Manufactured by Merck Group
Sourced in United States
The ZR-75-1 is a laboratory equipment product from Merck Group. It is designed to perform a specific function within a controlled laboratory environment. The details of its core function are presented in a factual and unbiased manner, without any interpretation or extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Mcf 7, by Merck Group (12 mentions)
Zr 75 1, by American Type Culture Collection (12 mentions)
Mcf 7, by American Type Culture Collection (11 mentions)
Mda mb 231, by American Type Culture Collection (6 mentions)
Mda mb 231, by Merck Group (5 mentions)
Skbr3, by American Type Culture Collection (4 mentions)
Bt474, by Merck Group (4 mentions)
Rpmi 1640 medium, by Merck Group (4 mentions)
Fetal bovine serum (fbs), by Merck Group (4 mentions)
Rpmi 1640, by Merck Group (3 mentions)
Sk br 3, by Merck Group (3 mentions)
L glutamine, by Thermo Fisher Scientific (3 mentions)
Bt474, by American Type Culture Collection (3 mentions)
Bovine insulin, by Merck Group (2 mentions)
Penicillin, by Merck Group (2 mentions)
Mda mb 468, by American Type Culture Collection (2 mentions)
L glutamine, by Merck Group (2 mentions)
Sodium pyruvate, by Merck Group (2 mentions)
Jimt 1, by Merck Group (2 mentions)
Dmem medium, by Merck Group (2 mentions)
16 protocols using zr 75 1
1
Breast Cancer Cell Line Culture and Signaling
2
Culturing Diverse Breast Cancer Cell Lines
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The human breast cancer cell lines MDA‐MB‐231, SKBR‐3, MDA‐MB‐468, ZR‐75‐1, BT474, MCF‐7 and T47D and normal breast epithelial cell line MCF‐10A were purchased from American Type Culture Collection (ATCC). MCF10A cells were cultured in DMEM/F12 (Sigma) supplemented with 5% horse serum (Thermo Fisher Scientific), 20 ng/mL EGF (BD Biosciences), 0.5 mg/mL hydrocortisone (Sigma‐Aldrich), 100 ng/mL cholera toxin (Sigma‐Aldrich), 10 mg/mL insulin (Gibco) and pen/strep. MDA‐MB‐231 cells were cultured in Leibovitz's L‐15 medium with 10% FBS at 37 1C without CO2. MDA‐MB‐436, ZR‐75‐1 and BT474 cells were cultured in RPMI‐1640 (Sigma) medium supplemented with 10% FBS. MCF‐7 were cultured in MEM (10% FBS, 1% NEAA, 0.01 mg/mL bovine insulin (Sigma‐Aldrich), 50 units/mL penicillin and 50 μg/mL streptomycin sulphate). T47D were cultured in RPMI medium (10% FBS, 1% NEAA, 50 units/mL penicillin and 50 μg/mL streptomycin sulphate).
3
Cell Culture of Human Cancer Cell Lines
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Human colon adenocarcinoma cell line HT-29 (Catalogue Number 91072201, ATCC HTB-38) and human breast carcinoma cell line ZR-75-1 (Catalogue Number 87012601, ATCC CRL 1500) were acquired from the European Collection of Authenticated Cell Cultures (ECCAC). In accordance with ECCAC instructions, HT-29 cells were cultured in McCoy’s 5a medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS) (Sigma Aldrich), 2 mM of L-Glutamine (Sigma-Aldrich), 100 U/ml of penicillin and 100 μg/ml of streptomycin (Sigma-Aldrich) at 37°C and 5% CO2. Cells were passaged when 80–90% confluence was reached, and the media was changed every 2–3 days. In accordance with ECCAC instructions, ZR-75-1 cells were cultured in RPMI 1640 (Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich), 1 mM sodium pyruvate (Sigma-Aldrich), 2 mM of L-Glutamine (Sigma-Aldrich) and 100 U/ml of penicillin and 100 μg/ml of streptomycin (Sigma-Aldrich) at 37°C and 5% CO2. Cells were passaged when 70–80% confluence was reached, and the media was changed every 2–3 days.
4
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).
5
Culturing Breast Cancer Cell Lines
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The human breast cancer cell lines MCF7, T47D, ZR-75-1 (ER+/HER2−), BT474 (ER+/HER2+), SKBR3 and JIMT-1 (ER−/HER2+) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). The MCF7, T47D, ZR-75-1, SKBR3 and JIMT-1 cell lines were cultured in an RPMI 1640 medium (Sigma, St. Louis, MO, USA), while the BT474 cell line was cultured in a DMEM medium (Sigma). All culture media were supplemented with 10% fetal bovine serum (FBS) (Sigma), an antibiotic-antimycotic solution (1×) (Sigma), and l -glutamine (2 mM) (Invitrogen GmbH, Karslruhe, Germany). The cultures were maintained in an incubator at 37 °C under 5% CO2.
6
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).
7
Culturing Diverse Breast Cell Lines
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All human breast cell lines used for this study were obtained from the ATCC (Rockville, MD, USA). The human breast cancer cell lines MDA-MB-231, MDA-361, MCF-7, ZR-75.1, BT-474, BT-20, T47D and BT-549 were routinely maintained in Dulbecco’s modified Eagle’s medium/Ham’s F12 (Sigma-Aldrich, Irvine, UK) supplemented with 10% foetal calf serum (FCS), and 1 × penicillin and streptomycin. MCF10A cells were maintained in MEGM (mammary epithelial growth medium) (Lonza, Slough, UK) supplemented with 100 ng ml−1 cholera toxin. The human microvascular endothelial cells (HMVECs) (Life Technologies, Paisley, UK) were maintained using Medium 131 supplemented with the addition of Microvascular Growth Supplement (MVGS) in flasks treated with Attachment Factor Protein (Life Technologies). All cells were incubated at 37 °C, with 5% CO2 and 95% humidity.
8
Culturing and Characterizing Breast Cancer Cell Lines
9
Cell Line Maintenance and Culture
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ZR-75-1, T47D, MCF7 and SKBR3 cell lines were purchased from the European Collection of Cell Cultures (Wiltshire, UK). All cell lines were maintained through continuous passaging, and were confirmed to be free of contamination by Mycoplasma spp. Cells were maintained in DMEM (ZR-75-1 cell line) or RPMI-1640 (other cell lines) media (Sigma-Aldrich, St. Louis, MO, USA), with 10% FBS (Sigma-Aldrich), 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco, Rockville, MD, USA) and 1.46 mg/mL L-glutamine (Gibco). Additionally, the growth medium for ZR-75-1 was supplemented with 1 nM β-estradiol (Sigma-Aldrich), and did not contain sodium pyruvate, which mediates elimination of H2O2 from the culture medium [21 (link)]. Tissue culture experimental techniques are further described in the Supplementary Methods (Additional file 1 ).
10
Cell Line Culture Protocol for Cancer Research
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Human embryonic kidney 293 (HEK293), human lung adenocarcinoma cell line A549, human ovarian adenocarcinoma cell lines SKOV-3 and OVCAR-3, human pancreatic carcinoma cell lines BxPC-3 and MIA-PaCa-2, human breast cancer cell lines AU-565, MCF-7 and ZR-75-1, human malignant mesothelioma cell lines H2052, H2452 and MSTO-211H, human prostate cancer cell line PC-3, were obtained from the American Type Culture Collection (ATCC; Manassas, VA). Human embryonic kidney 293A was obtained from Life Technologies (Carlsbad, CA, USA). Chinese hamster ovary (CHO) cells and CHO-hCAR cells stably expressing human CAR were provided by Jeffrey M. Bergelson [39] (link). All cells described above except AU-565, ZR-75-1, three kinds of malignant mesothelioma cell lines, CHO, and CHO-hCAR were cultured in Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 Ham, (DMEM/F12; Sigma-Aldrich, St. Louis, MO) containing 10% fetal bovine serum (FBS; Hyclone; Logan, UT), 2 mM L-glutamine, 100 U/ml penicillin, and 100 mg/ml streptomycin (Mediatech, Inc., Herndon, VA). The other cell lines were maintained in culture medium recommended by each supplier. All cells were incubated at 37°C in an atmosphere of 5% CO2 in air. Also, infected cells were maintained with medium containing 2% FBS, 2 mM L-glutamine, 100 U/ml penicillin, and 100 mg/ml streptomycin.
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