The cell lines used in this study (MCF10F, MCF10A and HEK293T) were purchased from ATCC, are originally derived from female tissue (Manassas, VA), and have been authenticated by shorten tandem repeat DNA profiling and validation that they are free of mycoplasma contamination. Cells were grown at 37°C with 5% CO2 and cultured in DMEM/F12 supplemented with 5% horse serum, 100 ng/ml cholera toxin, 5 μg/ml insulin, 0.5 μg/ml hydrocortisone, 50 ng/ml EGF, and 1% antibiotic/antimycotic (Corning, Corning, NY). Transfections were performed using FuGENE HD (Promega, Madison, WI). For lentivirus production, HEK293T cells were co-transfected with pCMV-VSV-G, pCMV-D8.2-Δvpr, and lentiviral expression vectors. For the generation of cell lines, MCF10F, MCF10A and HEK293T cells were incubated overnight in viral supernatants supplemented with 8 μl/ml protamine sulfate and subsequently selected for with antibiotics. Cell lines harboring multiple genetic manipulations were created by serial transductions. Clonal cell populations were created from single cells sorted into 96-well plates.
Mcf 10f
Manufactured by American Type Culture Collection
Sourced in United States
The MCF-10F is a cell line derived from normal human breast epithelium. It is used in research to study normal mammary gland biology and the early stages of breast cancer development.
Automatically generated - may contain errors
Lab products found in correlation
Mcf 7, by American Type Culture Collection (9 mentions)
Mda mb 231, by American Type Culture Collection (9 mentions)
Mcf 10a, by American Type Culture Collection (8 mentions)
Bt474, by American Type Culture Collection (6 mentions)
Hek293t, by American Type Culture Collection (5 mentions)
Skbr3, by American Type Culture Collection (5 mentions)
Hs578t, by American Type Culture Collection (4 mentions)
Mda mb 468, by American Type Culture Collection (4 mentions)
Hcc1937, by American Type Culture Collection (4 mentions)
Mda mb 453, by American Type Culture Collection (3 mentions)
Au565, by American Type Culture Collection (3 mentions)
Bt549, by American Type Culture Collection (3 mentions)
Mda mb 435, by American Type Culture Collection (3 mentions)
Antibiotic antimycotic, by Corning (2 mentions)
Fugene hd, by Promega (2 mentions)
Mda mb 361, by American Type Culture Collection (2 mentions)
Zr 75 1, by American Type Culture Collection (2 mentions)
Cal51, by American Type Culture Collection (2 mentions)
Hcc1143, by American Type Culture Collection (2 mentions)
Cama 1, by American Type Culture Collection (2 mentions)
15 protocols using mcf 10f
1
Cell Line Authentication and Genetic Manipulation
2
Culture of Breast Cancer Cell Lines
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Hs578T and MCF-10F cells were purchased from ATCC and cultured according to ATCC recommendations (ATCC, Manassas, VA). SUM149 cells were a generous gift from Dr. Stephen Ethier (Wayne State University, Detroit, MI). SUM149 cells were maintained in F-12 K Medium (Mediatech, Herndon, VA) containing 5 % FBS (Sigma-Aldrich), 0.5 μg/mL hydrocortisone (Sigma-Aldrich), 2 mM L-glutamine (Mediatech), 100 IU penicillin/100 μg/mL streptomycin (Mediatech), 10 μg/mL insulin (Sigma-Aldrich), and 5 μg/mL Plasmocin (Invivogen, San Diego, CA).
3
Cell Culture Protocol for Various Cell Lines
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Primary HMEC and human prostate epithelial cells (PHECs) were ordered from from Clonetics BioWhittaker (Walkersville, MD, USA). MCF-10F, RWPE1, U2OS, and 293T cells were ordered from ATCC. MGC-803 cells were obtained from Cell Resource Center, Chinese Academy of Medical Sciences, China. The HMEC/HMEC-hTERT and PHEC/PHEC-hTERT cells were maintained in serum-free mammary (Invitrogen) and prostate (PrEBM, Clonetics BioWhittaker) basal medium supplemented with growth factors, respectively. RWPE1 cells were cultured in keratinocyte serum-free medium (Invitrogen). U2OS and 293T cells were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS). MCF-10F cells were cultured in DMEM/F12 (1:1) media containing 5% horse serum (Invitrogen), 10 μg/mL insulin, 20 ng/mL epidermal growth factor, and 500 ng/mL hydrocortisone. MGC-803 cells were cultured in RPMI-1640 supplemented with 10% FBS. All cultures were kept at 37 °C with 5% CO2 atmosphere. Mycoplasma was tested as negative using Mycoplasma Detection Kit (Biotool, USA).
4
Breast Cancer Cell Lines Characterization
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ER- human breast non-tumorigenic epithelial cell lines (MCF-10A, MCF-10F and MCF-10-2A), ER+ human breast cancer cell lines (MCF-7 and T-47D) and ER- human breast cancer cell lines MDA-MB231 and SKBR-3) were from ATCC (Manassas, Virginia, USA). Tamoxifen was from Sigma-Aldrich. Cell culture medium (DMEM/F12), OPTI-MEM, Lipofectamine 2000 and TRIzol reagent were from Life Technologies (San Diego, CA, USA). Antibodies against β-actin and TFIIIC63 and c-Jun siRNA (Catalog No. SC-29224) were obtained from Santa Cruz Biotech (Santa Cruz, CA, USA). Mismatch RNA was described previously [24 (link)]. Histone H3 antibody were from Cell Signaling (Danvers, MA, USA). Brf1 antibody was from Bethyl laboratories Inc (Montgomery, TX, USA). The sequences of primers were described in (Supplements) [8 (link), 30 (link)].
5
Cell Line Authentication and Genetic Manipulation
6
Cytotoxicity and Antioxidant Evaluation of Ethanolic Extract and Compounds in MCF10F Cells
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The human breast non-tumorigenic cell line MCF10F (ATCC), considered the normal counterpart, were used to evaluate the cytotoxic effect and only MCF10F for antioxidant effect in vitro of ethanolic extract and pure compounds. These cells were cultured in specific media according to ATCC recommendations. The incubation conditions were established at 37 °C, complete humid atmosphere, 5% CO2 and 95% O2.
7
Relative Expression of SOX30 in Breast Cell Lines
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Human breast cancer cell lines including T47D, MDAMB231, MCF7, MDAMB453, BT474, and MDAMB468 were purchased from American Type Culture Collection (ATCC), and human normal breast cell line MCF‐10F was also purchased from ATCC. The T47D and BT474 cells were cultured in RPMI‐1640 medium (Gibco), and MDAMB231, MCF7, MDAMB453, MDAMB468, and MCF‐10F cells were cultured in DMEM medium (Gibco). All cells were maintained in the 95% air and 5% carbon dioxide (CO2). After culture, the relative expressions of SOX30 in these cells were determined by reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR) with MCF‐10F cells severed as control, which was performed according to a previous study.16
8
Breast Cancer Cell Line Cultivation
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The human BC cell lines MCF7 (ATCC HTB-22), ZR-75-1 (ATCC CRL-1599), BT-474 (ATCC HTB-20), BT-549 (ATCC HTB-122), MDA-MB-231 (ATCC CRM-HTB-26), AU565 (ATCC CRL-2351), and MDA-MB-361 (ATCC HTB-27) and the normal breast epithelial cell line MCF-10F (ATCC CRL-10318) were purchased from ATCC (ATCC, Manassas, VA, USA) and cultured in DMEM high-glucose medium [25 mM glucose and 4 mM glutamine, without pyruvate (Pyr), (Sigma Aldrich, St. Louis, MO, USA), promoting the same substrate availabilities for all cell lines. A description of the MCF7-TAMR, MCF7-rho0 and MCF7-Sph cells is provided in Appendix A .
9
Cell Line Authentication and Characterization
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The Research Resource Identifiers (RRIDs) for all cell lines used are provided in Table S2 . HMEC (human mammary epithelial cells) were a gift from M. R. Stampfer [39 ]. All other cell lines were purchased from DSMZ (“Deutsche Sammlung von Mikro-Organismen und Zellkulturen,” Braunschweig, Germany): CAL-51, HCC1143, HCC1937, and KPL-1, or ATCC (American Type Culture Collection, Manassas, USA): AU565, BT-474, CAMA-1, Hs 578T, Hs 578Bst, MCF-7, MCF-10A, MCF-10F, MDA-MB-231, MDA-MB-453, MDA-MB-468, SK-BR-3, T-47D, and ZR-75-1. Cell culture conditions of all cell lines were described previously [40 (link)]. DSMZ and ATCC authenticate all cell lines by STR profiling before distribution. Genomic DNA and total RNA were isolated from all cell lines immediately after receipt, i.e., within three to eight passages [40 (link),41 (link)].
10
Culturing Human Breast Cell Lines
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Human embryonic kidney HEK-293 T, Breast epithelial cell lines MCF-10A, MCF-10 F, MCF7, T47D and metastatic breast cancer cell lines MDA-MB-231, MDA-MB-435 and BT549 were purchased from American Type Culture Collection (ATCC) and preserved in our laboratory, MDA-MB-231-luc was a gift from Dr. Yongfeng Shang (Peking University Health Science Center). These cell lines were cultured in medium supplemented with 10 % FBS at 37 °C with 5 % CO2 in a humid atmosphere.
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