The cells (MDA-MB-231, NCCS, Pune) were grown in appropriate medium containing 10 % fetal bovine serum (Cat#092916854, M.P. Biochemicals) and 2 mM l -glutamine (Cat #12800-017, GIBCO) These were incubated at 37 °C, 5 % CO2, with 5000 cells in each well of the 96 well plate. The test drug was diluted and different concentrations were prepared with further incubation for 48 h. The addition of TCA (Cat #2844 M, Fisher Scientific) in 50 μl of cold 30 % (w/v) TCA (final concentration, 10 % TCA) for assay termination. The well washed and air dried sample is further treated with the Sulforhodamine B (SRB) (Cat# S9012 CAS- 3620, SIGMA) solution (50 μl) at 0.4 % (w/v) in 1 % acetic acid kept for 20 min at room temperature. Unbound dye recovered and residual dye washed off and treated with trizma base (Cat# T1378, SIGMA) before OD measured at 540 nm (Model# ELx808, BIOTEK instruments Inc). Percent growth was calculated on a plate-by-plate basis for test wells relative to control wells. Percent growth was expressed as the ratio of average absorbance of the test well to the average absorbance of the control wells × 100 [27 (link)].
Mda mb 231
Manufactured by MP Biomedicals
Sourced in India
MDA-MB-231 is a cell line derived from a human breast adenocarcinoma. It is commonly used in cancer research applications.
Automatically generated - may contain errors
Lab products found in correlation
Mcf 7, by MP Biomedicals (3 mentions)
Penicillin streptomycin, by MP Biomedicals (3 mentions)
Mcf 7, by National Centre for Cell Science (2 mentions)
Mda mb 231, by National Centre for Cell Science (2 mentions)
Elx808, by Agilent Technologies (1 mentions)
Sulforhodamine b srb, by Merck Group (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Trizma base, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by MP Biomedicals (1 mentions)
Mcf 10a, by American Type Culture Collection (1 mentions)
4 protocols using mda mb 231
1
SRB Cytotoxicity Assay for MDA-MB-231 Cells
2
Culturing Breast Cancer Cell Lines
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Breast cancer cell lines MCF7, T47D and MDA-MB-231 were purchased from National Centre for Cell Sciences (NCCS, Pune, India). MCF7 cells were cultured in Dulbecco′s Modified Eagle′s Medium (DMEM), whereas T47D and MDA-MB-231 cells in RPMI supplemented with 10% fetal bovine serum (FBS) and penicillin–streptomycin (MP Biomedicals, Bengaluru, India) at 37 °C, 5% CO2 and 95% humidity. MCF10A, a kind gift from Dr. Annapoorni Rangarajan (IISC, Bangalore, India), was maintained in DMEM F12 containing horse serum supplemented with hydrocortisone, EGF, insulin, cholera toxin and penicillin–streptomycin at 37 °C, 5% CO2 and 95% humidity. The cells were grown until 70–80% confluence and subcultured using Trypsin-EDTA.
3
Characterization of Breast Cancer Cell Lines
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The human breast cancer cell lines MCF-7 (RRID:CVCL_0031), ZR-75-1 (RRID:CVCL_0588), T-47D (RRID:CVCL_0553), SKBR3 (RRID:CVCL_0033), MDA-MB-231 (RRID:CVCL_0062), and MDA-MB-468 (RRID:CVCL_0419) were procured from the National Repository of Animal Cell Culture, NCCS Pune (Maharashtra, India). MCF10A (RRID:CVCL_0598) cell line was obtained from The American Type Culture Collection (ATCC), USA. All cell lines were screened for Mycoplasma and tested negative for it. They were independently verified by short tandem repeat (STR) DNA fingerprinting at the Institute of Life Sciences (Bhubaneswar, India). Experiments were conducted within five passages post-thawing. MCF-7, ZR-75-1, SKBR3, and MDA-MB-231 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM), whereas T-47D, MDA-MB-468 in Roswell Park Memorial Institute 1640 (RPMI) were supplemented with 10% fetal bovine serum (FBS) and 100 U/ml penicillin-streptomycin (MP Biomedicals, Bengaluru, India). MCF-10A was cultured in DMEM F12 containing horse serum (5%), hydrocortisone (0.5 mg/ml), EGF (20 ng/ml), insulin (10 μg/ml), cholera toxin (100 ng/ml), and 100 U/ml penicillin-streptomycin. Cells were maintained at 37°C, 5% CO2, and 95% humidity.
4
Breast Cancer Cell Line Culture
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Breast cancer cell lines MCF-7, T47D and MDA-MB-231 were purchased from National Centre for Cell Sciences (NCCS), Pune, India. The MCF-7 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) whereas T47D and MDA-MB-231 cells in Roswell Park Memorial Institute medium (RPMI) supplemented with 10% fetal bovine serum (FBS) and penicillin-streptomycin (MP Biomedicals) at 37 °C, 5% CO2 and 95% humidity. MCF10A, a kind gift from Dr. Annapoorni Rangarajan (IISC, Bangalore, India) was maintained in DMEM F12 containing horse serum supplemented with hydrocortisone, EGF, insulin, cholera toxin and penicillin-streptomycin at 37 °C, 5% CO2 and 95% humidity. The cells were grown until 70-80% confluence and then sub cultured with Trypsin-EDTA. All experiments involving treatment were performed in cells kept in phenol red free medium containing charcoal treated fetal bovine serum supplemented with penicillin-streptomycin for 48 h.
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