Mda mb 436

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MDA-MB-436 is a human breast cancer cell line derived from the pleural effusion of a patient with metastatic breast adenocarcinoma. It is commonly used in cancer research to study the biology and treatment of triple-negative breast cancer.

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Triple-negative MDA-MB-436 human breast adenocarcinoma cancer cells MDA-MB-436 cells

241 protocols using mda mb 436

Breast Cancer Cell Line Characterization

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MCF-7 (ER+/PR+/HER2−, BRCA1 proficient), MDA-MB-231 (ER−/PR−/HER2−, BRCA1 proficient), MDA-MB-468 (ER−/PR−/HER2−, BRCA1 proficient) and MDA-MB-436 (ER−/PR−/HER2−, BRCA1 deficient) were purchased from ATCC and were grown in RPMI (MCF-7) or DMEM (MDA-MB-231, MDA-MB-468 and MDA-MB-436) medium with the addition of 10% foetal bovine serum and 1% penicillin/streptomycin. Cells in culture were routinely checked for mycoplasma contamination by PCR (Sigma, catalog no. MP0035).The cells characterisation were performed by ATCC and passaged in the laboratory for fewer than 6 months.

Arora A., Parvathaneni S., Aleskandarany M.A., Agarwal D., Ali R., Abdel-Fatah T., Green A.R., Ball G.R., Rakha E.A., Ellis I.O., Sharma S, & Madhusudan S. (2016). Clinicopathological and functional significance of RECQL1 helicase in sporadic breast cancers. Molecular cancer therapeutics, 16(1), 239-250.

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Cell Culture Conditions for TNBC Lines

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The epithelial TNBC cell lines MDA-MB-231 (ATCC HTB-26™; Manassas, VA, USA) and MDA-MB-436 (ATCC HTB-130™; Manassas, VA, USA), and the normal breast epithelial cell line MCF-10A (ATCC CRL-10317™; Manassas, VA, USA), were purchased from the American Type and Culture Collection (ATCC; Manassas, VA, USA). The MDA-MB-231 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (ATCC; Manassas, VA, USA) supplemented with 10% fetal bovine serum (HyClone; Logan, UT, USA) and 1% antibiotic (100 IU/mL of penicillin and 100 µg/mL of streptomycin; HyClone; Logan, UT, USA), and incubated at 37 °C, with a humidified atmosphere of 5% CO₂. The MDA-MB-436 cells were cultivated in Leibovitz’s L-15 medium (ATCC; Manassas, VA, USA) supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin solution (100 IU/mL of penicillin and 100 µg/mL of streptomycin; ATCC; Manassas, VA, USA), 10 µg/mL of insulin (Gibco; Life Technologies; Grand Island, NY, USA), and 16 µg/mL of glutathione (Sigma-Aldrich; St. Louis, MO, USA), and maintained at 37 °C, with a humidified atmosphere of 0% CO₂. The MCF-10A cells were cultured in DMEM/F12 medium supplemented with a MEGM Kit (Lonza Pharma and Biotech; Basel, Switzerland) and incubated at 37 °C, with a humidified atmosphere of 5% CO₂.

Mohammadhosseinpour S., Ho L.C., Fang L., Xu J, & Medina-Bolivar F. (2022). Arachidin-1, a Prenylated Stilbenoid from Peanut, Induces Apoptosis in Triple-Negative Breast Cancer Cells. International Journal of Molecular Sciences, 23(3), 1139.

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Cell Culture Protocols for Cancer and Control

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Human normal breast epithelial cell line (MCF-10A), human TNBC cell lines (MDA-MB-231, MDA-MB-436, BT-549, HCC1937) as well as human embryonic kidney cell line (HEK-293 T) were all procured from the American Type Culture Collection (ATCC; Manassas, VA, USA) and preserved at 37 °C with 5% CO₂. Leibovitz’s L-15 medium (Thomas Scientific, Swedesboro, NJ, USA) was applied to incubate MDA-MB-231 and MDA-MB-436 cells; ATCC-formulated RPMI-1640 medium (ATCC) was used for cultivating BT-549 and HCC1937 cells; Mammary Epithelial Cell Growth Medium (MEGM; Gibco, Grand Island, NY, USA) was adopted for cultivating MCF-10A cells; Dulbecco’s modified Eagle medium (DMEM; Gibco) was used for incubating HEK-293 T cells. 10% fetal bovine serum (FBS; Gibco) together with 1% penicillin-streptomycin (Sigma-Aldrich, St. Louis, MO, USA) was also applied to all media.

Qian J., Lei X., Sun Y., Zheng L., Li J., Zhang S., Zhang L., Li W., Shi J., Jia W, & Tang T. (2021). Long non-coding RNA SNHG8 enhances triple-negative breast cancer cell proliferation and migration by regulating the miR-335-5p/PYGO2 axis. Biology Direct, 16, 13.

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Cell lines for BRCA2 and SIRT studies

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DLD1 BRCA2^+/+ and ^–/– (Horizon Discovery), HAP1 WT, SIRT1^–/–, SIRT3^–/–, SIRT6^–/– (Horizon Discovery), CAL51 (DSMZ), HEK293T, MDA-MB-436 (ATCC) and U2OS WT and U2OS HPF1^–/– (gift from Ivan Ahel) cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS (with 10 µg/ml insulin in the case of MDA-MB-436). SUM149 cells (Asterand Bioscience) were maintained in Ham’s F-12 medium supplemented with 5% FBS, 10 µg/ml insulin and 1 µg/ml hydrocortisone.
All human cell line identities were confirmed by STR typing and verified free of mycoplasma infection using Lonza MycoAlert.

Bajrami I., Walker C., Krastev D.B., Weekes D., Song F., Wicks A.J., Alexander J., Haider S., Brough R., Pettitt S.J., Tutt A.N, & Lord C.J. (2021). Sirtuin inhibition is synthetic lethal with BRCA1 or BRCA2 deficiency. Communications Biology, 4, 1270.

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TNBC Cell Line Culture and DRAIR Overexpression

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Human TNBC cell lines MDA-MB-436 and SUM149PT from ATCC were cultured in DMEM medium (Cat # 11,054,001, Thermo Fisher Scientific, Shanghai, China) supplemented with 10% FBS (Cat # MFCD00132239, Sigma-Aldrich, Shanghai, China), streptomycin (10 μg/ml, Cat # 3810–74-0, Sigma-Aldrich, Shanghai, China) and penicillin (100 U/ml, Cat # 87–08-1, Sigma-Aldrich, Shanghai, China) in an incubator at 37°C with 95% humidity and 5% CO₂.
MDA-MB-436 (ATCC, USA) and SUM149PT (ATCC, USA) cells were overexpressed with DRAIR by transfecting cells with DRAIR expression vector (pcDNA3.1) using Neon Electroporation Transfection device (Thermo Fisher Scientific, Shanghai, China). Transected cells were cultured in fresh media prior to subsequent assays.

Wang P., Peng W, & Peng J. (2022). Diabetes regulated anti-inflammatory lncRNA is overexpressed in triple-negative breast cancer and predicts chemoresistance and tumor recurrence. Bioengineered, 13(5), 12718-12725.

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Characterization of Mammary Tumor Cell Lines

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The 545-GFP cell line was derived from Brca1^Co/Co MMTV-Cre (Brca1-MSK) mice, which develop only primary mammary tumors and have minimal metastatic ability, as determined by labeling with GFP. The 628-GFP cell line was also derived from Brca1-MSK mice, which not only develop primary mammary tumors but also develop multiple metastatic tumors in other organs, which were identified by the GFP label. B477 is a Brca1-WT mammary gland epithelial cell line, and G600 was derived from Brca1-MT mammary gland epithelial cells. T47D (catalog HTB-133), MDA-MB-231 (catalog CRM-HTB-26), MDA-MB-436 (catalog HTB-130), and HEK293T (catalog ACS-4500) cells were purchased from American Type Culture Collection (ATCC). All the cells above were cultured with high-glucose DMEM containing 10% FBS and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin; Gibco). All cell lines were cultured at 37°C in an atmosphere with 5% CO₂. All other human breast cancer cell lines, including MDA-MB-436, MDA-MB-468, T47D, and HEK293T, were purchased from ATCC.

Xu J., Su S.M., Zhang X., Chan U.I., Adhav R., Shu X., Liu J., Li J., Mo L., Wang Y., An T., Lei J.H., Miao K., Deng C.X, & Xu X. (2022). ATP11B inhibits breast cancer metastasis in a mouse model by suppressing externalization of nonapoptotic phosphatidylserine. The Journal of Clinical Investigation, 132(5), e149473.

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Cisplatin Effects on TNBC Cell Lines

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The normal human breast epithelial cell line MCF10A and the human TNBC cell lines HCC1937, BT-549, MDA-MB-231, MDA-MB-436, and MDA-MB-468 were purchased from ATCC (Manassas, VA, USA). MCF10A cells were cultured in DMEM/F12 medium supplemented with 5% heat-inactivated horse serum, 0.5 mg/mL hydrocortisone, 20 ng/mL EGF, 10 μg/mL insulin, 100 ng/mL cholera toxin, 100 IU/mL penicillin, and 100 μg/mL streptomycin. HCC1937 cells were maintained in RPMI 1640 medium (Life Technologies, Carlsbad, CA, USA). BT-549 cells were cultured in RPMI 1640 medium with 10% fetal bovine serum (FBS) and 10 μg/mL insulin. MDA-MB-231 and MDA-MB-468 cells were cultured in ATCC-formulated Leibovitz’s L-15 medium supplemented with 10% FBS, while MDA-MB-436 cells were grown in the same medium with 10 μg/mL insulin and 16 μg/mL glutathione. All the cells were maintained in a humidified incubator at 37°C with 5% CO₂. Cells were passaged by trypsinization after reaching approximately 80%–90% confluence. The detached cells were seeded into a new flask at a density of 2.0 × 10³ cells/cm².
For determination of the effect of cisplatin on TNBC cells, cells were cultured with or without 10, 20, 50, 100, and 200 μM DDP for 24 h or with 10 μM DDP for 24 h and then collected for subsequent experiments.

Ma H.Y., Li Y., Yin H.Z., Yin H., Qu Y.Y, & Xu Q.Y. (2020). TNFAIP8 Promotes Cisplatin Chemoresistance in Triple-Negative Breast Cancer by Repressing p53-Mediated miR-205-5p Expression. Molecular Therapy. Nucleic Acids, 22, 640-656.

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Cell Line Authentication and Testing

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HCC70, HCC1143, HCC1187, HCC1937, HCC1954, HCC1806, AU565, BT474, MCF7, MDAMB231, MDAMB436 and BT549 cell lines were purchased directly from ATCC. The MB468 cell line was engineered to express luciferase and was obtained from Dr. A. Kung (Memorial Sloan Kettering Cancer Center). All cell lines were tested for Mycoplasma infection by PCR every 3 months. Only early-passage cell lines were used and cells were kept in culture no longer than 21 days. Cell lines were obtained in 2012 with the exception of MDAMB231 that was obtained in 2016. Cell lines were authenticated by short tandem repeat (STR) analysis. Bortezomib was purchased from LC Laboratories. E7107 (18 (link)) was kindly provided by H3 Biomedicine, Inc.

Chan S., Sridhar P., Kirchner R., Lock Y.J., Herbert Z., Buonamici S., Smith P., Lieberman J, & Petrocca F. (2017). Basal-A Triple Negative Breast Cancer Cells Selectively Rely on RNA Splicing for Survival. Molecular cancer therapeutics, 16(12), 2849-2861.

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Basal-Like Breast Cancer Cell Line Cultures

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A total of 8 different human basal-like breast cancer cell lines (MDA-MB231, MDA-MB468, MDA-MB436, BT20, BT549, HCC70, HCC1937, and HS.578t) were obtained from ATCC and cultured as recommended by ATCC. Six SUM series of basal like breast cancer cell lines (SUM149, SUM102, SUM159, SUM1315, SUM225 and SUM229) were provided by Professor Stephen P. Either from the Medical University of South Carolina. SUM159, SUM149 and SUM 225 were cultured in Ham’s F12 medium with 5% FBS with supplementary of 5 µg/ml insulin and 1 µg/ml hydrocortisone (5% IH), SUM 1315 cells were cultured in F12 medium with 5% FBS with supplementary of 5 µg/ml insulin and 10 ng/ml EGF (5% IE). SUM102 cells were cultured in F-12 medium with supplementary of serum replacement factors (5 mM ethanolamine, 10 mM HEPEs, 5 µg/ml transferrin, 10 nM triiodothyronine, 50 µM sodium selenite, and 0.5 g/L BSA), 5 µg/ml insulin, 1 µg/ml hydrocortisone and 10 ng/ml EGF (SFIHE). The mouse breast cancer cell line 4T1 was obtained from ATCC and was cultured in high glucose DMEM and supplemented by 10% FBS, NEAA and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin).

Wu L., Meng F., Dong L., Block C.J., Mitchell A.V., Wu J., Jang H., Chen W., Polin L., Yang Q., Dou Q.P, & Wu G. (2019). Disulfiram and BKM120 in Combination with Chemotherapy Impede Tumor Progression and Delay Tumor Recurrence in Tumor Initiating Cell-Rich TNBC. Scientific Reports, 9, 236.

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Breast Cancer Cell Line Cultivation

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Breast cancer cell lines HS578T (#HTB-126) and MDAMB436 (#HTB-130) were obtained from ATCC, and EAC cells OE19 were obtained from Sigma-Aldrich, and were authenticated by the STR-PCR analysis. All cell lines used in this study were negative for mycoplasma using in-house tests. The HS578T cell line was cultured in Dulbecco’s modified Eagle medium (DMEM) (Thermo Fisher Scientific, USA) and MDAMB436 were cultured in L-15 medium with 10 µg/ml insulin (Thermo Fisher Scientific, USA) and 16 µg/ml glutathione (iCell Bioscience Inc, Shanghai). The EAC cell line OE19 was cultured in RPMI-1640 medium (Thermo Fisher Scientific, USA). All the medium was supplemented with 10% fetal bovine serum (FBS) (Omega Scientific, USA) and 100 U/ml penicillin and 100 mg/ml streptomycin (Thermo Fisher Scientific, USA).

Zheng Y., Huang G., Silva T.C., Yang Q., Jiang Y.Y., Koeffler H.P., Lin D.C, & Berman B.P. (2021). A pan-cancer analysis of CpG Island gene regulation reveals extensive plasticity within Polycomb target genes. Nature Communications, 12, 2485.

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