Human breast cancer cell lines MDA361, T47D, SKBR3, MDA-MB-468 (MDA-468), MCF-12A, and MCF-10A were purchased from American Type Culture Collection (ATCC). MDA361, T47D, MDA-468, and SKBR3 were cultured in DMEM/F-12 (Mediatech Inc.) supplemented with 10% FBS and penicillin/streptomycin. HMLE (kindly provided by Dr. R. A. Weinberg) cell lines were cultured in 1:1 Dulbecco’s Modified Eagle’s Medium (DMEM)/Ham’s F-12 medium (Mediatech Inc.) supplemented with 5% FBS (Clontech), 100 units/ml penicillin-streptomycin (Invitrogen), 2 mml-glutamine (Invitrogen), 10 ng/ml human epidermal growth factor (EGF) (Invitrogen), 0.5 μg/ml hydrocortisone (Sigma), and 10 μg/ml insulin (Sigma).MCF-12A and MCF-10Awere cultured in 1:1 Dulbecco’s Modified Eagle’s Medium (DMEM)/Ham’s F-12 medium (Mediatech Inc.) supplemented with 5% Horse serum (Clontech), 100 units/ml penicillin-streptomycin, 10 ng/ml human epidermal growth factor (EGF), 0.5 μg/ml hydrocortisone, and 10 μg/ml insulin .
Mcf 12a
Manufactured by Merck Group
Sourced in United States
MCF-12A is a cell line derived from normal human mammary epithelial cells. It is commonly used in cell culture research related to breast biology and breast cancer studies.
Automatically generated - may contain errors
Lab products found in correlation
Hydrocortisone, by Merck Group (4 mentions)
Mcf 12a, by American Type Culture Collection (4 mentions)
Glutamine, by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Insulin, by Merck Group (3 mentions)
Mcf 7, by American Type Culture Collection (3 mentions)
Epidermal growth factor (egf), by Merck Group (3 mentions)
Mda mb 231, by American Type Culture Collection (3 mentions)
Hydrocortisone, by Takara Bio (2 mentions)
Insulin, by Takara Bio (2 mentions)
Penicillin streptomycin, by Takara Bio (2 mentions)
Fetal bovine serum (fbs), by Takara Bio (2 mentions)
Horse serum, by Takara Bio (2 mentions)
Human epidermal growth factor egf, by Takara Bio (2 mentions)
Human epidermal growth factor, by Thermo Fisher Scientific (2 mentions)
Dmem ham s f 12 medium, by Corning (2 mentions)
Mcf 10a, by American Type Culture Collection (2 mentions)
Skbr3, by American Type Culture Collection (2 mentions)
Mda 361, by American Type Culture Collection (2 mentions)
Mcf 7, by Thermo Fisher Scientific (2 mentions)
7 protocols using mcf 12a
1
Culture and Maintenance of Breast Cancer Cell Lines
2
Cell Lines and Culture Conditions
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The breast cancer tumorigenic cell lines MDA-MB-231 and MCF-7, the breast normal-like cell lines MCF12A and MCF10A, HEK293, and Jurkat cells were obtained from ATCC (Rockville, MD). MDA-MB-231, MCF-7 and HEK293 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies Grand Island, NY) supplemented with 10% fetal bovine serum and antibiotics. Jurkat cells were maintained in RPMI (Life Technologies) supplemented with 10% fetal bovine serum and antibiotics. The normal cell lines MCF12A and MCF10A were cultured in a 1:1 mixture of Ham’s F12 and DMEM supplemented with 10% bovine insulin, 20ng/ml epidermal growth factor and 500ng/ml hydrocortisone (Sigma, St Louis, MO). TTP/zfp36+/+ and TTP/zfp36−/− mouse embryonic fibroblasts were obtained as described previously (21 (link)) and were grown in DMEM. HEK293 Tet-On Advanced cells (Clontech, Mountain View, CA) were used in tetracycline-induced expression experiments and were cultured in DMEM supplemented with 10% Tet System Approved FBS (Clontech), 100 μg/ml G418 (Sigma) and 5% Penicillin-Streptomycin (Invitrogen, Carlsbad, CA). All transfections were carried out in reduced serum media using Lipofectamine 2000 (Invitrogen).
3
Breast Cancer Tissue Collection and Cell Culture
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This study was approved by the Ethics Committee of Jinshan Hospital. A total of 127 pairs of paraffin-embedded breast IDC tissues and adjacent normal tissues, as well as 105 benign fibroadenomas, were consecutively collected from 2012 to 2016 for IHC study. The follow-up of patients was from January, 2012 to January, 2019. Fresh breast tissues of normal, benign fibroadenomas, and IDC were used for mRNA and protein extraction. All tissues from patients without neoadjuvant therapies such as hormonal therapy, chemotherapy or radiotherapy were immediately frozen in liquid nitrogen after surgery and stored at -80°C for subsequent use. Human non-cancerous cell line MCF-12A and cancerous cell lines MCF-7 and MDA-MB-231 were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). MCF-12A cells were cultured in DMEM/F12 (Sigma-Aldrich, Cat# D84378, Saint Louis, MO, USA) with 5% FBS (HAO YANG BIOLOGICAL, Cat# TBD15HT, Tianjin, China), 10 μg/ml insulin (Wanbang Pharma, Cat#H19994040, Xuzhou, China), 500 ng/ml hydrocortisone (Shenggong, Cat# A610506, Shanghai, China), and 10 ng/ml EGF (Sigma-Aldrich, Cat# E9644). MCF-7 and MDA-MB-231 cells were cultured in RPMI1640 (Sigma-Aldrich, Cat# R8758) with 10% FBS. The cells were incubated and maintained in a humidified atmosphere of 5% CO2 at 37°C.
4
Breast Cancer Cell Line Characterization
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Human immortalized, untransformed mammary epithelial cell line, MCF-12A, and human breast cancer cell lines, MCF-7 and MDA-MB-231, were obtained from American Type Culture Collection (Rockville, MD, USA). MCF-7 is estrogen receptor (ER) and progesterone receptor (PR) positive, but MDA-MB-231 is ER and PR negative. All cell lines were maintained in antibiotic-free media at 5%CO2 and 37°C. MCF-7 and MDA-MB-231 were grown in alpha minimum essential medium (a-MEM) supplemented with 10% fetal calf serum and 1 mM glutamine (GIBCO Invitrogen, Carlsbad, CA, USA). Immortalized, untransformed MCF-12A cells were cultured in a 1:1 mixture of Ham’s F12 medium and Dulbecco’s Modified Eagle’s Medium containing 0.1 μg/mL cholera enterotoxin, 10 μg/mL insulin, 0.5 μg/mL hydrocortisone, 20 μg/mL epidermal growth factor, and 5% horse serum (Sigma Chemical Co., St. Louis, MO, USA). Cells were detached from tissue culture flasks by digestion with 0.05% trypsin and 0.53 mM EDTA.
5
Culturing Human Breast Cancer and Normal Cells
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MCF-7 human breast cancer cells and MCF-12A human noncancerous breast epithelial cells were obtained from the American Type Culture Collection (ATCC). MCF-7 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Sigma, USA) supplemented with 10% Fetal Bovine Serum (FBS) (Biological Industries, Israel) and 1% penicillin/streptomycin (Biowest, USA). MCF-12A cells were maintained in DMEM Ham’s F12 Medium (Sigma, USA), which contains 10% FBS, 1% penicillin/streptomycin, 10 μg/ml insulin, 20 ng/ml EGF and 0.5 mg/ml hydrocortisone. The cells were incubated at 37 °C in a humidified atmosphere containing 5% CO2.
6
Culture and Maintenance of Breast Cancer Cell Lines
7
Establishment of Stable Breast Cell Lines
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The immortalized normal mammary epithelial cell line, MCF12A, and the luminal breast cancer cell line, MCF7, as well as 293T cells, were purchased from American Type Culture Collection (ATCC). MCF12A cells were grown in DMEM-F12 medium supplemented with 5% horse serum, epidermal growth factor (20 ng/ml), insulin (10 ng/ml), cholera toxin (100 ng/ml), hydrocortisone (500 ng/ml) and gentamycin (Sigma). MCF7 cells were cultured with DMEM medium supplemented with 10% FBS, penicillin (50 U/ml), and streptomycin (50 U/ml). To establish stable cell lines, shRNA plasmids (targeting human or mouse FOXP1, Sigma) or CRISPR plasmids (pLV-CRISPR-hTET2, Vector Builder) were co-transfected with lentiviral packaging plasmids (pPAX2 and pMD2.G) into 293T cells. After 48 h incubation, lentiviruses were harvested and used to infect the target cells. Stable cell lines were selected with puromycin (2 μg/mL) treatment for 7 days.
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