MCF-7, MDA-MB-231, ZR-75-1, MDA-MB-453 and T47D cells were acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were cultured in DMEM or RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 mg/mL streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.) at 37◦C and 5%CO2 in humidified air. The normal breast cell line MCF-10A was purchased from the Institute of Biochemistry and Cell Biology of the Chinese Academy of Sciences (Shanghai, China), The MCF-10A cells were cultured in DMEM/F12, with 10% FBS, 20 ng/mL EGF, 0.1 mg/mL CT, 10 mg/mL insulin, and 500 ng/mL hydrocortisone (Sigma). The transfection was performed using the lipofectamine 3000 (Invitrogen, Carlsbad, CA), followed by the protocol recommended by the manufacturer. The plasmids used were synthesized by GenePharma (Shanghai, China). Knockdown of FOXN4 and P53 were achieved by using lentivirus carrying the relative shRNA sequences (Sigma). The establish confirmation of stable lentiviral transduction was determined using infection with shSCR as negative controls.
Mcf 10a
Manufactured by Merck Group
Sourced in United States, China
MCF-10A is a human breast epithelial cell line. It is non-tumorigenic and is commonly used as a model for normal breast epithelial cells in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Mcf 10a, by American Type Culture Collection (27 mentions)
Cholera toxin, by Merck Group (23 mentions)
Insulin, by Merck Group (21 mentions)
Mda mb 231, by American Type Culture Collection (21 mentions)
Hydrocortisone, by Merck Group (20 mentions)
Mcf 7, by American Type Culture Collection (20 mentions)
Epidermal growth factor (egf), by Merck Group (15 mentions)
Mcf 7, by Merck Group (9 mentions)
Dmem f12, by Merck Group (9 mentions)
Penicillin, by Merck Group (8 mentions)
Mda mb 468, by American Type Culture Collection (8 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions)
Streptomycin, by Merck Group (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Horse serum, by Thermo Fisher Scientific (7 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (6 mentions)
Mcf 7, by Thermo Fisher Scientific (6 mentions)
Mda mb 231, by Merck Group (5 mentions)
Non essential amino acids (neaa), by Merck Group (5 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
58 protocols using mcf 10a
1
Cell Lines Establishment and Transfection
2
Cell Culture Protocols for Multiple Cell Lines
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NRK (normal rat kidney fibroblasts), HeLa (human epithelial), MDA-MB-231 (human breast adenocarcinoma), MCF 10A (human fibrocystic disease), and MCF7 (human breast adenocarcinoma) cells were obtained from the American Type Culture Collection (Manassas, VA). NRK, HeLa and MCF7 cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; Life Technologies). MCF 10A and MDA-MB-231 cells were maintained in DMEM-F12 (Life Technologies). For all cell lines the media were supplemented with 10% heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin (Life Technologies), and 5 μg/ml plasmocin (InvivoGen, San Diego, CA). MCF 10A and MCF7 media were further supplemented with 10 μg/ml insulin (Sigma-Aldrich). MCF 10A medium was also supplemented with 100 ng/ml cholera toxin, 20 ng/ml EGF, and 0.5 μg/ml hydrocortisone (Sigma-Aldrich).
3
Isogenic Cell Lines for E-cadherin Knockout
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MCF10A and MCF10A CDH1−/− isogenic cell lines were purchased from Sigma-Aldrich (#CLLS1042, Sigma-Aldrich, St Louis, MO, USA). Cells were cultured in Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 with GlutaMAX (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 5% horse serum (Invitrogen, Carlsbad, CA, USA), 20 ng/mL EGF (Peprotech, Rehovot, Israel), 100 ng/mL cholera toxin (Sigma-Aldrich, St Louis, MO, USA), 0.5 µg/mL hydrocortisone (Sigma-Aldrich, St Louis, MO, USA) and 10 µg/mL insulin (Novo Nordisk Pharmaceuticals Ltd., Bagsværd, Denmark).
The NCI-N87 cell line was purchased from ATCC, and the NCI-N87 CDH1−/− cell line was generated within our laboratory [13 (link)]. Cells were grown in Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 with GlutaMAX (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA).
All cells were cultured in a 37 °C humidified incubator at 5% CO2.
The NCI-N87 cell line was purchased from ATCC, and the NCI-N87 CDH1−/− cell line was generated within our laboratory [13 (link)]. Cells were grown in Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 with GlutaMAX (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA).
All cells were cultured in a 37 °C humidified incubator at 5% CO2.
4
Culturing Hepatoma and Colon Cancer Cells
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Hepatoma cell lines HepG2, colon cancer cell line SW480 were obtained from Cell Lines Bank, Chinese Academy of Science (Shanghai, China). The cells were maintained in DMEM containing 10% fetal bovine serum, 100 U/ml of penicillin and 100 mg/ml streptomycin sulfate and incubated at 37°C in a humidified atmosphere of 5% CO2. MCF-10A and rictor-null MCF-10A cells were purchased from Sigma, and culture in DMEM/F12 medium containing 5% horse serum, 2.5 mM L-glutamine, 10 mg/mL human insulin, 0.5 mg/mL hydrocortisone, 10 ng/mL EGF, and 100 ng/mL cholera toxin, 100 U/ml of penicillin and 100 mg/ml streptomycin sulfate.
5
Culturing Breast Cancer and Normal Cell Lines
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BC cells (HCC70 and MDA-MB-231) and human mammary gland epithelial cells (MCF-10A) were bought from the American Type Culture Collection (Manassas, VA, USA). These cells were cultured in mammary epithelial cell growth medium (MEGM) BulletKit (Lonza, Basel, Switzerland) (MCF-10A cells) or Roswell Park Memorial Institute-1640 medium (Sigma, St Louis, MO, USA) (HCC70 and MDA-MB-231) supplemented with 10% fetal bovine serum (FBS) (Sigma) and 1% streptomycin/penicillin (Solarbio, Beijing, China) and maintained at 37 °C in a moist atmosphere with 5% CO2.
6
Culturing Breast Cancer and Normal Epithelial Cells
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The human BC cell line MDA-MB-468 and the immortalized normal breast epithelial cell line MCF-10A were purchased from the American Type Culture Collection. BC cells MCF-7, MDA-MB-231 and MDA-MB-453 were obtained from the Cell Bank of Chinese Academy of Sciences. MDA-MB-231, MDA-MB-453 and MDA-MB-468 cells were cultured in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (HyClone; GE Healthcare Life Sciences). MCF-7 cells were cultured in Dulbecco's Modified Eagle's medium (Gibco; Thermo Fisher Scientific, Inc.) with 10 µg/ml human recombinant insulin (Sigma-Aldrich; Merck KGaA) and 10% fetal bovine serum (HyClone; GE Healthcare Life Sciences). MCF-10A cells were cultured in RPMI-1640 medium with 10% fetal bovine serum and 100 ng/ml cholera toxin (Sigma-Aldrich; Merck KGaA). Cells were cultured at 37°C in a humidified atmosphere containing 5% CO2.
7
Culturing Diverse Breast Cancer Cell Lines
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The human breast cancer cell lines MDA‐MB‐231, SKBR‐3, MDA‐MB‐468, ZR‐75‐1, BT474, MCF‐7 and T47D and normal breast epithelial cell line MCF‐10A were purchased from American Type Culture Collection (ATCC). MCF10A cells were cultured in DMEM/F12 (Sigma) supplemented with 5% horse serum (Thermo Fisher Scientific), 20 ng/mL EGF (BD Biosciences), 0.5 mg/mL hydrocortisone (Sigma‐Aldrich), 100 ng/mL cholera toxin (Sigma‐Aldrich), 10 mg/mL insulin (Gibco) and pen/strep. MDA‐MB‐231 cells were cultured in Leibovitz's L‐15 medium with 10% FBS at 37 1C without CO2. MDA‐MB‐436, ZR‐75‐1 and BT474 cells were cultured in RPMI‐1640 (Sigma) medium supplemented with 10% FBS. MCF‐7 were cultured in MEM (10% FBS, 1% NEAA, 0.01 mg/mL bovine insulin (Sigma‐Aldrich), 50 units/mL penicillin and 50 μg/mL streptomycin sulphate). T47D were cultured in RPMI medium (10% FBS, 1% NEAA, 50 units/mL penicillin and 50 μg/mL streptomycin sulphate).
8
Culturing Breast Cancer Cell Lines
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Human breast cancer cell lines, such as MDA-MB-231 and MCF-7, and the human mammary epithelial cell line MCF-10A were purchased from ATCC-LGC-Promochem (Wesel, Germany). These cancer cell lines were cultured in 4.5 g/l DMEM with added 10% Fetal Calf Serum and 1% penicillin-streptomycin (Biochrom, Berlin, Germany) in an incubator at 37°C, 5% CO2 and 95% humidity. Instead, the mammary epithelial cell line MCF-10A was cultured in a 1:1 mixture of DMEM (4.5 g/l glucose, L -glutamine) and Ham’s F12 medium with added 5% Horse Serum, 1 g/l Cholera toxin stock solution, 10 mg/ml Insulin (Sigma-Aldrich, I9278), 1 g/l Hydrocortison stock solution, 100 μg/ml epidermal growth factor (EGF) and 1% penicillin/streptomycin 100× (P/S) under the same conditions as mentioned above. For all experiments, cells with passage numbers of 5–25 at about 80% confluency were harvested.
9
Cell Line Cultivation Protocols
10
Culturing Breast Cancer Cell Lines
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MDA-MB-231, T-47D and MCF-7 human breast cancer cell lines and MCF-10A normal human breast epithelial immortalized cell line were obtained from American Type Culture Collection (ATCC) (Rockville, USA). MDA-MB-231, T-47D and MCF -7 breast cancer cells were cultured in DMEM supplemented with 10 % (v/v) heat inactivated FBS, 2 mM L-glutamine, 100 U/ml of penicillin, and 100 μg/ml of streptomycin. MCF-10A cells were maintained in DMEM-F12 (Sigma-Aldrich, USA) supplemented with 10 % (v/v) FBS, 100 U/ml of penicillin, 100 μg/ml of streptomycin, 2 mM of L-glutamine, 20 ng/ml of epidermal growth factor (Sigma-Aldrich, USA), 10 μg/ml of insulin (Sigma-Aldrich, USA), 100 ng/ml of cholera toxin (Sigma-Aldrich, USA) and 1 μg/ml of hydrocortisone (Sigma-Aldrich, USA). Cells were cultured in 75 cm2 culture flasks at 37 °C under humid environment in an incubator having 5 % CO2.
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