MCF7 (ATCC® HTB-22™), T47D (ATCC® HTB-133™) and MDA-MB-231 (ATCC® HTB-26™) cell lines were purchased from LGC standards. MCF7, T47D, MDA-MB-231 were grown and maintained in respectively DMEM /Ham F12, RPMI-1640 and DMEM media (DUBELCCO). Media were supplemented with 10% fetal bovine serum (FBS), 1% glutamine (Gibco) and antibiotics (Penicillin/Streptomycin), and cells grown at 37°C in a 5% CO2 humidified atmosphere. For defined estrogen stimulation culture experiments, cells at 70% confluence were trypsinized and plated for 12 hours, washed twice and a steroid depleted media (phenol red-free DMEM/ham F12 supplemented with 2.5% charcoal stripped calf bovine serum- PAA) was added. Cells were cultured for at least 72 hours before treatment with 17β-Estradiol (E2) (Tocris bioscience) 10 nM, ICI 182,780 100 nM (Tocris bioscience) or vehicle control (ethanol (Sigma Aldrich) 0.1%).
Htb 133
Manufactured by American Type Culture Collection
Sourced in United States
HTB-133 is a human prostate carcinoma cell line. It is a well-established and widely used model for prostate cancer research. The HTB-133 cell line is available from the American Type Culture Collection (ATCC) for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Htb 22, by American Type Culture Collection (7 mentions)
Htb 26, by American Type Culture Collection (5 mentions)
Mcf 7, by American Type Culture Collection (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Mda mb 231, by American Type Culture Collection (3 mentions)
Dmem f12, by Thermo Fisher Scientific (2 mentions)
Mcf 10a, by American Type Culture Collection (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Ethanol, by Merck Group (1 mentions)
Glutamine, by Thermo Fisher Scientific (1 mentions)
17β estradiol e2, by Bio-Techne (1 mentions)
Ccl 185, by American Type Culture Collection (1 mentions)
Crl 1740, by American Type Culture Collection (1 mentions)
Pcs 440 010, by American Type Culture Collection (1 mentions)
Pcs 600 010, by American Type Culture Collection (1 mentions)
Skbr3, by American Type Culture Collection (1 mentions)
Mcf 10a, by Thermo Fisher Scientific (1 mentions)
Bt549, by American Type Culture Collection (1 mentions)
Bt 549, by Thermo Fisher Scientific (1 mentions)
Sk br 3, by Thermo Fisher Scientific (1 mentions)
16 protocols using htb 133
1
Estrogen-Responsive Breast Cancer Cell Lines
2
Culturing Multiple Cell Lines
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Normal human bronchial epithelial cells (ATCC® PCS-300-010™, VA, USA), prostate epithelial cells (ATCC PCS-440-010™), mammary epithelial cells (ATCC PCS-600-010™), A549 (human lung tumor cells, ATCC CCL-185™), LNCaP (human prostate adenocarcinoma cells, ATCC CRL-1740™) and T-47D (human breast tumor cells, ATCC HTB133™) were used. Cells were cultured in appropriate cell growth kit, supplemented with additional growth factors as provided by ATCC, and maintained in a 5% CO2 humidified incubator at 37°C.
3
Breast Cancer Cell Culture Protocols
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In vitro studies used luminal (T-47D, ATCC HTB-133), triple-negative (BT-549 ATCC HTB-122), and HER2 (SK-BR-3, ATCC CRL-10317) cells, and an immortalized mammary epithelial cell line (MCF 10A, ATCC HTB-30). Human embryonic kidney 293T cells were used for the lentivector experiments. BT-549 and T-47D were purchased from ATCC, whereas MCF 10A and 293T were kindly donated by Dr. Cos (University of Cantabria) and Dr. Escors (Navarrabiomed, FMS), respectively. T-47D, BT-549 and SK-BR-3 were cultured in RPMI-1640, supplemented with 1% penicillin/streptomycin and 10% fetal bovine serum (FBS) (Invitrogen, Life Technologies, Carlsbad, CA, USA). MCF-10A was cultured in DMEM/F-12 (Invitrogen, Life Technologies) supplemented with 1% penicillin/streptomycin, 10% FBS, 20 ng/mL of epidermal growth factor (EGF), 500 ng/mL of hydrocortisone and 10 μg/mL of insulin (Sigma-Aldrich, St Louis, MO, USA). 293T cells were cultured in DMEM supplemented with 1% penicillin/streptomycin and 10% FBS (Invitrogen, Life Technologies). Cells were grown in a humidified atmosphere (5% CO2, 37°C). Cells were always passaged before they reached confluence.
4
Estrogen Depletion and Tamoxifen Treatment
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The human BCa cell lines MCF-7 and T47D were purchased from American Type Culture Collection (ATCC® HTB-22TM and ATCC® HTB-133 respectively, Manassas, VA, USA).
To deplete estrogen, cells were cultured in phenol red-free RPMI 1640 containing 2.5% HyClone Charcoal/Dextran-Treated Fetal Bovine Serum (SH30068.03, Thermo Scientific, Waltham, MA, USA) for 24 h. Next, 17-β estradiol (E2, Sigma-Aldrich, St. Louis, MO, USA) in ethanol was added to the culture medium at a final concentration of 0, 0.1, 1, or 10 nM, and the cells were cultured for another 24 h. Tamoxifen was purchased from Sigma and added to the culture medium at a final concentration of 2 μM.
To deplete estrogen, cells were cultured in phenol red-free RPMI 1640 containing 2.5% HyClone Charcoal/Dextran-Treated Fetal Bovine Serum (SH30068.03, Thermo Scientific, Waltham, MA, USA) for 24 h. Next, 17-β estradiol (E2, Sigma-Aldrich, St. Louis, MO, USA) in ethanol was added to the culture medium at a final concentration of 0, 0.1, 1, or 10 nM, and the cells were cultured for another 24 h. Tamoxifen was purchased from Sigma and added to the culture medium at a final concentration of 2 μM.
5
Cell Culture Protocols for Cancer Cell Lines
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The human melanoma cells lines 501mel, MM200, and WM293a (kindly donated by Professor Dorothy Bennet, St George's University of London) were cultured in Roswell Park Memorial Institute Medium (RPMI)-1640 (Sigma Aldrich, USA). The human breast adenocarcinoma cell lines MCF-7 (ATCC® HTB-22) and T47D (ATCC® HTB-133) were cultured in Dulbecco's Modified Eagle medium:nutrient mixture F-12 (Ham) (DMEM/F-12) (Gibco by Thermo Fisher Scientific, USA). The human embryonal rhabdomyosarcoma cell line RD (ATCC® CCL-136) and the human alveolar rhabdomyosarcoma cell line RH30 (kindly provided by Professor Judith Davie, Southern Illinois University) were cultured in DMEM (Sigma Aldrich, USA). All culture medium was supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin. Cells were maintained at 37°C in a 5% CO2 humidified incubator and medium was replaced every 2–3 days.
6
Culturing Mammary Cell Lines
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MCF-7 and T-47D cell lines were purchased from ATCC (#HTB22 and #HTB-133, respectively), and Bre-80 was kindly provided by Roger Reddel (CMRI, Sydney, Australia). Cell lines were stored in liquid nitrogen vapour phase with Mycoplasma testing and short tandem repeat profiling performed for cell authentication prior to storage. The immortalised mammary epithelial Bre-80 cell line was cultured as previously described [24 (link)]. The breast cancer cell lines MCF-7 and T-47D were cultured in RPMI 1640 medium supplemented with fetal calf serum, penicillin/streptomycin, and 10 μg/ml insulin. All cell lines were cultured at 37 °C in a humidified 5% CO2 atmosphere.
7
Cell Line Characterization and Culture
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HEK293T, MCF7, and T47D cell lines were purchased from ATCC (CRL-3216, HTB-22, and HTB-133, respectively). Ishikawa cells were purchased from Millipore Sigma (99040201). MCF7:WS8 cells were kindly donated by Dr. Clodia Osipo. MCF7 Y537S ESR1 breast cancer cells were kindly donated by Dr. Sarat Chandarlapaty. All cell lines were tested for mycoplasma quarterly and their identities confirmed using STR profiling through ATCC before the start of experiments. WT and Y537S ESR1 MCF7 and MCF7:WS8 cells were cultured in DMEM supplemented with 10% FBS and 6.5 μg/mL bovine insulin. T47D cells were cultured with RPMI supplemented with 10% FBS. Ishikawa cells were cultured with MEM supplemented with 2 mM l -glutamine, 1% non-essential amino acids, and 5% FBS.
8
Comprehensive Cell Culture Protocol for Cancer Research
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In this project, the colorectal cancer cell line HCT116 (ATCC® CCL-247™), androgen-dependent prostate cancer cell line PC-3 (ATCC® CRL-1435™), breast cancer cell lines T47D (ATCC® HTB-133™), MCF7 (ATCC® HTB-22™) and finally, epithelial embryonic kidney cell line HEK293 (ATCC® CRL-1573™) were used. All media were completed with %10 fetal bovine serum (FBS) and %1 penicillin streptomycin amphotericin (PSA). All cells were cultured periodically at 37 °C, 5% (v/v) CO2 and 95% (v/v) air in T-75 flasks. Different media were used for the different cell lines based on ATCC standards: MCF7 and HEK293 cell lines were cultured in DMEM High Mmedium (ATCC, 30-2003); T47D and PC-3 cell lines were cultured in RPM 1640 medium (ATCC, 30-2001); HCT116 cell line was cultured in DMEM F12 medium (ATCC, 30-2006). RPM 1640 medium was further completed with 0.2 unit/mL bovine insulin for the T47D cell line. Cells were passaged when they reached %80 confluency in order to support proliferation.
9
Breast Cancer Cell Line Cultivation
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Breast cancer cell lines MCF-7 (American Type Culture Collection [ATCC]® HTB-22™), T-47D (ATCC® HTB-133™), BT-474 (ATCC® HTB-20™), and MDA-MB-231 (ATCC® HTB-26™) were purchased from ATCC (Manassas, VA, USA). MCF-7 cells were cultivated in DMEM (F0475) with 10% FBS and 2 mM l -glutamine; T-47D cells in RPMI-1640 with 10% FBS, 0.1 units/mL human insulin, and 2 mM l -glutamine; BT-474 cells in DMEM (F0445) with 8% FBS, 12% Panexin, and 2 mM l -glutamine; and MDA-MB-231 cells in DMEM (FG0445) with 10% FBS, 1% MEM NEAA, and 2 mM l -glutamine. All the cells were cultured at 37°C with 5% CO2. Passaging was performed according to the manufacturer’s instructions.
10
Culturing Breast Cancer Cell Lines
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MDA-MB-231 cells (ATCC® HTB-26™) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Corning) and supplemented with 10% fetal bovine serum (FBS) (Gemini) and 1% Penicillin-Streptomycin (P/S) (Corning). T47D cells (ATCC® HTB-133™) were transfected with a lentiviral plasmid to express CCT2 with a FLAG tag (DYKDDDDK) hereafter referred to as T47D-CCT2, as previously described [13 (link)]. T47D cells were cultured in Roswell Park Memorial Institute Medium (RPMI-1640) (Corning) supplemented with 10% FBS (Gemini), 1% P/S (Corning), and 0.2 units/mL human recombinant insulin (Santa Cruz). 0.5 μg/mL puromycin dihydrochloride (ThermoFisher) was added to maintain plasmids.
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