Hec1A, AGS, and HCC1500 cells were acquired from ATCC. All other cell lines (HCC1806, SUM149, BT549, HCC1937, HCC70, BT20, MCF7, MDAMB453, EFM19, ZR75-1, and T47D) were acquired from the Brugge Lab at Harvard Medical School, and were authenticated by STR analysis. All cell lines were of female origin. Cell lines were regularly tested for mycoplasma using the MycoAlert Mycoplasma Detection Kit (Lonza). Cells were grown in human plasma-like medium according to the published formulation (Cantor et al., 2017 (link)) with 5% dialyzed FBS (Sigma) and Pen/Strep (Invitrogen) at 37 degrees C with 5% CO2. Media was changed at least every two days. As needed, cells were incubated in RPMI media (R9010-01, US Biological Life Sciences) with or without serine and glycine with 5% dialyzed FBS and Pen/Strep. Cells were counted using a Z1 Coulter Particle Counter (Beckman Coulter) and growth rates were calculated using the following formula: growth rate = ln(final cell number/initial cell number)/time.
Hcc1500
Manufactured by American Type Culture Collection
Sourced in United States
The HCC1500 is a high-capacity cell culture incubator designed for efficient cell growth. It features precise temperature and CO2 control to provide an optimal environment for cell cultivation. The HCC1500 offers a large interior chamber with multiple shelves to accommodate a variety of cell culture vessels.
Automatically generated - may contain errors
Lab products found in correlation
Mcf 7, by American Type Culture Collection (9 mentions)
Bt549, by American Type Culture Collection (8 mentions)
Hcc1937, by American Type Culture Collection (8 mentions)
Mda mb 231, by American Type Culture Collection (8 mentions)
Mda mb 468, by American Type Culture Collection (7 mentions)
Skbr3, by American Type Culture Collection (6 mentions)
Hcc38, by American Type Culture Collection (6 mentions)
Zr 75 1, by American Type Culture Collection (6 mentions)
Hcc1954, by American Type Culture Collection (6 mentions)
Hcc70, by American Type Culture Collection (6 mentions)
Mda mb 436, by American Type Culture Collection (6 mentions)
Bt 20, by American Type Culture Collection (6 mentions)
Bt474, by American Type Culture Collection (5 mentions)
Hcc1569, by American Type Culture Collection (5 mentions)
Mda mb 157, by American Type Culture Collection (5 mentions)
Hs578t, by American Type Culture Collection (4 mentions)
Mda mb 453, by American Type Culture Collection (4 mentions)
Au565, by American Type Culture Collection (4 mentions)
Hcc1806, by American Type Culture Collection (4 mentions)
Hcc1143, by American Type Culture Collection (3 mentions)
20 protocols using hcc1500
1
Culture of Breast Cancer Cell Lines
2
Breast and Kidney Cell Assays
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Authenticated MCF7, T47D, MCF10A, HCC1500, and 293T cells were purchased from ATCC and used for experiments within the first 10 passages. Cultures were checked for Mycoplasma every 6 months (Lonza) and cells maintained at 37°C and 5% CO2 in RPMI or DMEM + 10% FBS and 1% Pen-Strep. GDC-0941, BYL719, GDC-0032, and GDC-0068 were purchased from Selleck, and MI-136, MI-503, and MM-102 were purchased from Cayman. siRNAs (Silencer Select) were purchased from Invitrogen (Negative control #1, 4390843; siKMT2D, s528766).
3
Breast cancer cell line culture
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The cell lines used in this study: AU565, BT20, BT474, BT483, BT549, HCC1143, HCC1500, HCC1569, HCC1937, HCC1954, HCC38, HCC70, Hs578T, MDA-MB-134, MDA-MB-175, MDA-MB-361, MDA-MB-436, MDA-MB-453, MDA-MB-468, SKBR3, and ZR751 were obtained from ATCC, Rockville, MD, USA. MCF-7 cells were obtained from Michigan Cancer Foundation, Detroit, MI, USA. The cell lines HBL100, MDA-MB-157, MDA-MB-231 and T47D were obtained from EG&G Mason Research Institute, Worcester, MA, USA.
The cell lines AU565, BT20, BT474, BT549, HBL100, Hs578T, MCF-7, MDA-MB-134, MDA-MB-157, MDA-MB-175, MDA-MB-231, MDA-MB-361, MDA-MB-436, MDA-MB-453, MDA-MB-468, SKBR3, T47D and ZR571 were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 6 mM L-glutamine, 20 mM HEPES and 10 μg/ml human insulin (CSL-Novo, North Rocks, NSW, Australia). The remaining cell lines were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated FBS, 6 mM L-glutamine, 1 mM sodium pyruvate and 20 mM HEPES. The MYC over-expressing MCF7 cells have been previously described
[11 (link),27 (link)] and were cultured in the same conditions as the parental cells.
The CDK4/6 inhibitor PD0332991 was purchased from Selleck Chemicals (Houston, TX, USA), CDK2 inhibitor SNS-032 from Symansis (Auckland, New Zealand) and CDK1 inhibitors, RO-3306 and CGP74514A, from Calbiochem (San Diego, CA, USA).
The cell lines AU565, BT20, BT474, BT549, HBL100, Hs578T, MCF-7, MDA-MB-134, MDA-MB-157, MDA-MB-175, MDA-MB-231, MDA-MB-361, MDA-MB-436, MDA-MB-453, MDA-MB-468, SKBR3, T47D and ZR571 were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 6 mM L-glutamine, 20 mM HEPES and 10 μg/ml human insulin (CSL-Novo, North Rocks, NSW, Australia). The remaining cell lines were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated FBS, 6 mM L-glutamine, 1 mM sodium pyruvate and 20 mM HEPES. The MYC over-expressing MCF7 cells have been previously described
[11 (link),27 (link)] and were cultured in the same conditions as the parental cells.
The CDK4/6 inhibitor PD0332991 was purchased from Selleck Chemicals (Houston, TX, USA), CDK2 inhibitor SNS-032 from Symansis (Auckland, New Zealand) and CDK1 inhibitors, RO-3306 and CGP74514A, from Calbiochem (San Diego, CA, USA).
4
Culture of Breast Cancer Cell Lines
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Hec1A, AGS, and HCC1500 cells were acquired from ATCC. All other cell lines (HCC1806, SUM149, BT549, HCC1937, HCC70, BT20, MCF7, MDAMB453, EFM19, ZR75-1, and T47D) were acquired from the Brugge Lab at Harvard Medical School, and were authenticated by STR analysis. All cell lines were of female origin. Cell lines were regularly tested for mycoplasma using the MycoAlert Mycoplasma Detection Kit (Lonza). Cells were grown in human plasma-like medium according to the published formulation (Cantor et al., 2017 (link)) with 5% dialyzed FBS (Sigma) and Pen/Strep (Invitrogen) at 37 degrees C with 5% CO2. Media was changed at least every two days. As needed, cells were incubated in RPMI media (R9010-01, US Biological Life Sciences) with or without serine and glycine with 5% dialyzed FBS and Pen/Strep. Cells were counted using a Z1 Coulter Particle Counter (Beckman Coulter) and growth rates were calculated using the following formula: growth rate = ln(final cell number/initial cell number)/time.
5
Characterization of Breast Cancer Cell Lines
6
Authentication and Culturing of Human Breast Cancer Cell Lines
7
Breast Cancer Cell Line Cultures
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The BT549, BT20, DU4475, HCC38, HCC70, HCC1500, HCC1569, HCC1954, HCC1806, HCC1143, HCC1937, HS578T, MDA-MB-157, MDA-MB-436, MDA-MB-453, MDA-MB-231, and MDA-MB-468 cell lines were purchased from the American Type Culture Collection; CAL51, CAL148, and CAL851 cells were obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen; SUM185PE and SUM149PT cells were purchased from Asterand Bioscience; MFM223 cells were purchased from Sigma-Aldrich; and CAL120 cells were a gift from Professor Elgene Lim from the Garvan Institute of Medical Research, Darlinghurst, NSW, Australia. All the above cell lines were cultured in RPMI supplemented with 10% FBS and 0.023 IU/ml (or 10 µg/ml) insulin. MDA-MB231(LM2) cells (a kind gift from Dr. Joan Massague, Sloan-Kettering Memorial Institute, New York, NY; Minn et al., 2005 (link)) were cultured in DMEM supplemented with 10% FBS.
8
Culturing Breast Cancer Cell Lines
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MCF7, SKBR3, HCC1500, ZR-75-1 (all EpCAMpos) and MDA-MB-231 (EpCAMlow/neg) breast cancer cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, US). TMX2-28 cells (EpCAMpos) were a generous gift from Prof. K.F. Arcaro (University of Massachusetts, MA, US) [31 (link)]. All cell lines were cultured in RPMI 1640 containing 10% (v/v) fetal calf serum and 1% (v/v) penicillin/streptomycin (all Gibco/Life Technologies, Darmstadt, Germany). SKBR3 and ZR-75-1 cells were cultured without any supplements, whereas culture media for MCF7 and TMX2-28 were supplemented with 25 mM HEPES. MDA-MB-231 cells were maintained with supplementation of 2 mM L-glutamine and 20 mM HEPES; for HCC1500 cells 2 mM L-glutamine, 1 mM sodium pyruvate (all Gibco/Life Technologies) and 0.45% (v/v) D-(+) glucose solution (Sigma-Aldrich, Munich, Germany) were added. All cells were cultured at 37°C in a humidified atmosphere with 5% CO2. The same culture conditions were fulfilled for cells grown on coverslips in order to check for marker expression by immunofluorescence staining. At approx. 80% confluence, cells were washed once with PBS (Life Technologies) and fixed with ice cold methanol. Until further use, coverslips were stored at -20°C.
9
Culturing Human Mammary and Breast Cancer Cells
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Human mammary epithelial (HMEC) cells were purchased from Lonza (Basel, Switzerland) and breast carcinoma cell lines 184A1, AU-565, CAMA-1, DU-4475, HCC-1500, HCC-1569, HCC-1806, HCC-1937, HCC-202, HCC-70, Hs578T, MCF-7, MDA-MB-157, MDA-MB-175V11, MDA-MB-468, T-47D, UACC-3199 and ZR-75-30 were purchased from American Type Culture Collection (Manassas, VA). Cells were tested negative for mycoplasma contamination and validated for species and unique DNA profile using short tandem repeat analysis by the provider or by the authors. All cell lines were cultured in RPMI Medium 1640 (Life Technologies, Carlsbad, CA) supplemented with 10% fetal bovine serum, 1% Antibiotic-Antimycotic containing penicillin, streptomycin and Fungizone (Invitrogen, Carlsbad, CA), and 1% HEPES at 37 °C in an atmosphere containing 5% CO2.
10
Coculture of NK-92 and Breast Cancer Cells
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All human breast tissue derived cell lines (hTERT-HME1, PMEC, BT20, BT474, BT549, HCC1500, HCC1954, MCF-7, MDA-MB-231, SKBR3 and T47D), the leukemia cell line K562 and NK-92 cell line were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in the conditions recommended by ATCC. When the effector (NK-92) and target (breast tissue derived cell lines and K562) cells were cocultured together for assays, the effector complete medium without IL-2 (i.e. Alpha Minimum Essential medium without ribonucleosides and deoxyribonucleosides but with 2 mM L-glutamine and 1.5 g/L sodium bicarbonate supplemented with 12.5% horse serum, 12.5% fetal bovine serum, 0.2 mM inositol, 0.1 mM 2-mercaptoethanol, 0.02 mM folic acid, 100 units/ml penicillin and 100 µg/ml streptomycin) was used. 16–24 hours before effector-target coculture, NK-92 cells were cultured at 500,000 cells/mL in fresh effector complete medium containing 300 U/mL recombinant human IL-2 (PeproTech, Hamburg, Germany). All other media, serum and supplements were from Life Technologies (ThermoFisher Scientific, Waltham, MA, USA), Sigma-Aldrich (St. Louis, MO, USA) or ATCC.
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