The patient‐derived IBC cell line SUM‐149 (BioIVT, Westbury, NY, USA) was cultured in Ham’s F12 medium (Life Technologies, Carlsbad, CA, USA) with 10% fetal bovine serum (FBS), as previously described [54 (link), 55 (link), 56 (link)]. SUM‐149 IBC cells were seeded 1 : 3 in 10‐cm plates. Once they reached 70% confluency, 3 mL of culture media was collected every 72 h from three plates, while regular media served as control. For total protein extraction, the cells were harvested with a cell scraper and lysed on ice using NP‐40 lysis buffer containing one protease inhibitor tablet (Roche Diagnostics Corporation, Indianapolis, IN, USA). Cells were incubated for 10 min on ice and vortexed intermittently. Supernatants were collected after centrifugation [21,256 g (RCF), 4 °C, 10 min], and protein concentration was quantified using the Precision Red protein assay kit (Cytoskeleton, Inc., Denver, CO, USA), as described previously [54 (link), 56 (link), 57 ].
Sum149
Manufactured by BioIVT
Sourced in United States, United Kingdom
SUM149 is a cell line derived from triple-negative breast cancer. It is a useful model for studying this aggressive form of breast cancer.
Automatically generated - may contain errors
Lab products found in correlation
Bt474, by American Type Culture Collection (2 mentions)
Mcf 10a, by American Type Culture Collection (2 mentions)
Mda mb 231, by American Type Culture Collection (2 mentions)
Mcf 7, by American Type Culture Collection (2 mentions)
Mda mb 468, by American Type Culture Collection (2 mentions)
Sum159, by BioIVT (2 mentions)
Pen strep, by Thermo Fisher Scientific (2 mentions)
F12 medium, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Insulin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions)
Humec ready medium, by Thermo Fisher Scientific (2 mentions)
Ham s f12 medium, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor tablet, by Roche (1 mentions)
Precision red protein assay kit, by Cytoskeleton (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Mda mb 231, by LGC (1 mentions)
Mcf 7, by LGC (1 mentions)
Skbr3, by LGC (1 mentions)
7 protocols using sum149
1
Culturing Patient-Derived IBC Cell Line
2
Comprehensive Cell Line Database
3
Comprehensive Cell Line Database
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293T, HeLa, MDA-MB-231, MDA-MB-468, HMEC, MCF7, MCF10A and BT474 cells were all obtained from the American Type Culture Collection (ATCC). SUM149 and SUM159 cells were obtained from BioIVT (formerly Astrand). All cell lines are female.
4
Cell Culture Maintenance Protocol
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Cell lines were all obtained from the American Type Culture Collection, or BioIVT. 293T, HeLa, MDA-MB-231, MDA-MB-468, HMEC, MCF7, MCF10A, SUM159, and BT474 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; GIBCO) supplemented with 10% FBS (Atlanta Biologicals) and 1% Pen/Strep (GIBCO). SUM149 cells were obtained from BioIVT and cultured in either F-12 medium (GIBCO) supplemented with 5% FBS (Atlanta Biologicals), 10mM HEPES pH7.5, 1μg/mL hydrocortisone, 5μg/mL insulin (GIBCO) and 1% Pen/Strep (GIBCO) or in HuMEC Ready Medium (GIBCO). All cells were incubated at 37°C and 5% CO2.
5
Retinoic Acid Treatment in TNBC Cell Lines
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All cell lines were obtained from the American Type Culture Collection (ATCC), with the exception of SUM149 cells that were obtained from BioIVT (previously Asterand), were cultured as per the supplier’s recommendations. The TNBC patient-derived xenograft (PDX) 7482 was obtained as a low-passage cryopreserved tumor piece from Dr. Michael Lewis (Baylor College of Medicine). Prior to experimentation, the cryopreserved PDX tumor pieces were revived and surgically implanted in the mammary fat pad of a NOD/SCID female mouse for expansion for 5 weeks. PDX 7482 originated from a grade 3, stage 2 primary tumor, breast carcinoma [32 (link)]. For retinoic acid treatment, cell lines were treated with 100 nM all-trans retinoic acid (ATRA; Sigma–Aldrich) for 24 h, and then collected for quantitative polymerase chain reaction (QPCR) analysis as described below.
6
Cell Culture Maintenance Protocol
7
Culturing of Human Breast Cancer Cell Lines
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Three non-IBC human breast cancer cell lines MDA-MB-231 (ATCC® HTB-26™), MCF7 (ATCC® HTB-22™), SKBR3 (ATCC® HTB-30™) from LGC Promochem (Wesel, Germany), and one IBC cell line SUM149 (BIOIVT, West Sussex, UK) were used in this study. All cell lines were cultured in DMEM medium with 10% fetal bovine serum and 1% of penicillin/streptomycin antibiotic mixture, except SUM149 cells that were cultured in HAM’s F12 medium with 5% fetal bovine serum, 5 mM HEPES, 1 µg/mL hydrocortisone, 5 µg/mL insulin and 1% of penicillin/streptomycin antibiotic mixture. All cell lines were incubated at 37 °C in 5% CO2. Cells were detached at 80% of the confluence with 0.5% trypsin/EDTA (Invitrogen, Illkirch, France). Cells in suspension were centrifuged at 420 g for 3 min, then pellets were resuspended. Cell viability was assessed by trypan blue exclusion assay.
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