Hs578t

Manufactured by American Type Culture Collection

Sourced in United States, France, Japan

The Hs578T is a human breast carcinoma cell line. It is a well-established in vitro model for the study of breast cancer.

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433 protocols using hs578t

TNBC Cell Line Culture Protocol

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The human TNBC cell lines Hs578T and MDA-MB-231 were obtained from the American Type Culture Collection and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with fetal bovine serum (FBS) (10% v/v), insulin (Hs578T cell only), and penicillin G/streptomycin 1% (v/v) at 37 °C under 5% CO₂. The absence of culture contamination by Mycoplasma species was confirmed before the experiments.

Uddin M.J., Russo D., Haque M.A., Çiçek S.S., Sönnichsen F.D., Milella L, & Zidorn C. (2021). Bioactive Abietane-Type Diterpenoid Glycosides from Leaves of Clerodendrum infortunatum (Lamiaceae). Molecules, 26(14), 4121.

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3D Culture of Breast Cancer Cells

Cited 2 times

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HMT-3522, S1, and T4-2 cells (a kind gift from Dr. Mina J Bissell) were maintained as previously described (29 (link)). MDA-MB-231 and BT549 (ATCC) cells were cultured in DMEM/F12 (Sigma), Hs578T in DMEM (Sigma), both supplemented with 10% fetal bovine serum, 10 units/ml of penicillin and 0.1 mg/ml of streptomycin (Invitrogen). In 3D culture, cells were plated on Matrigel^® and maintained in the culture medium containing 5% Matrigel. T4 cells were seeded at a density of 2.1×10⁴ cells per cm²; while MDA-MB-231 and Hs578T (ATCC) were seeded at 1.4×10⁴ cells per cm². For production of lentivirus, 293FT cells were transfected with shRNA vector plus packaging vectors using FuGENE (Promega). Culture supernatants containing viral particles were collected 48h after transfection. Cells were infected with lentivirus and selected using puromycin for at least three days. Cell migration and invasion assay were performed as previously described (30 (link)).

Zhu J., Xiong G., Fu H., Evers B.M., Zhou B.P, & Xu R. (2015). Chaperone Hsp47 drives malignant growth and invasion by modulating an ECM gene network. Cancer research, 75(8), 1580-1591.

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Culturing Triple-Negative Breast Cancer Cell Lines

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Human TNBC cell lines (HCC38, BT-549, HS578T, MDA-MB231, BT-20) were purchased from ATCC (Virginia, USA) maintained in DMEM/ RPMI-1640 (Life Technologies, CA, USA) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37°C jacketed with 5% CO₂ atmosphere according to ATCC recommendations.

Pearlman A., Rahman M.T., Upadhyay K., Loke J, & Ostrer H. (2019). Ectopic Otoconin 90 expression in triple negative breast cancer cell lines is associated with metastasis functions. PLoS ONE, 14(2), e0211737.

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Breast Cancer Cell Line Characterization

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MCF7, BT549, MDAMB231, MDAMB468, Hs578T, SUM159PT, BT549, and BT20 breast cancer cell lines were obtained from ATCC (Manassas, VA, USA). MCF7 cells were grown in DMEM medium, the other cell lines were grown in RPMI medium, with 10% fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 µg/mL). Cells were plated 24 h before treatment with different drugs at the indicated concentrations. Recombinant human EGF, heregulin (HRG), IGF1, angiotensin II (Ang II), and angiotensin 1–7 (Ang 1–7) were obtained from Sigma Chemical Co (St. Louis, MO, USA). Recombinant HGF and FGF were from Abcam. Recombinant PDGF-BB was from Gibco. Trastuzumab, gifitinib, lapatinib, sunitinib, OSI-906, and doxorubicin were obtained from Selleck Chemicals. The PRCP inhibitor (PRCP-7414, referred to as PRCPi in text) was from Calbiochem (catalog number 504044).

Duan L., Calhoun S., Perez R.E., Macias V., Mir F., Pergande M.R., Gattuso P., Borgia J.A, & Maki C.G. (2022). Prolyl Carboxypeptidase Maintains Receptor Tyrosine Kinase Signaling and Is a Potential Therapeutic Target in Triple Negative Breast Cancer. Cancers, 14(3), 739.

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Breast Cancer Cell Line Maintenance

Cited 3 times

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MDA-MB-231, BT-20, HCC1937 breast cancer cells and HMEC were maintained and grown as previously described [14 (link), 15 (link), 31 (link)]. MDA-MB-453, MDA-MB-157, Hs578T were purchased in the year 2015 from ATCC and grown according to the ATCC recommended culture conditions. All cell lines were authenticated by genomic STR profile.

Singha P.K., Pandeswara S., Geng H., Lan R., Venkatachalam M.A., Dobi A., Srivastava S, & Saikumar P. (2019). Increased Smad3 and reduced Smad2 levels mediate the functional switch of TGF-β from growth suppressor to growth and metastasis promoter through TMEPAI/PMEPA1 in triple negative breast cancer. Genes & Cancer, 10(5-6), 134-149.

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Cell Culture Protocols for Cancer Cell Lines

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HEK293T, BT-549 and Hs578T cells were obtained from ATCC. BT549 was maintained in RPMI 1640 medium (Gibco) supplemented with 10% FBS. HEK293T cells were cultured in Dulbecco’s modified Eagle medium (DMEM; Gibco) supplemented with 10% FBS. Hs578T cells were maintained in DMEM supplemented with 10% FBS and 10 μg/ml insulin. All cell lines obtained from cell banks listed above are tested for authentication using short tandem repeat profiling and passaged for fewer than 6 months, and routinely assayed for mycoplasma contamination.

Maryam A, & Chin Y.R. (2021). ANLN Enhances Triple-Negative Breast Cancer Stemness Through TWIST1 and BMP2 and Promotes its Spheroid Growth. Frontiers in Molecular Biosciences, 8, 700973.

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Cell Culture Conditions for Metabolic Studies

Cited 1 time

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H1299 (CRL-5803), 786-O (CRL-1932), A498(HTB-44), H226 (CRL-5826), A549(CRL-7909), T98G(CRL-1690), Hs578T(HTB-126), and HEK293T (CRL-3216) cell lines were obtained from ATCC. UMRC6 (#08090513) was purchased from Sigma. All cell lines used in this study were free of mycoplasma contamination (per testing done by the vendor). All the cells were cultured in a 37 °C incubator in a 5% CO₂ atmosphere. H1299 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and 10,000 U/mL penicillin-streptomycin. 786-O, A498, UMRC6, H226, A549, T98G, Hs578T, and HEK293T cells were cultured in Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum and 10,000 U/mL of penicillin-streptomycin. For the glucose deprivation experiments, cells were cultured in glucose-free DMEM (Life Technologies #11966025) supplemented with dialyzed fetal bovine serum as described previously^{34 (link),35 (link)}.

Yan Y., Teng H., Hang Q., Kondiparthi L., Lei G., Horbath A., Liu X., Mao C., Wu S., Zhuang L., James You M., Poyurovsky M.V., Ma L., Olszewski K, & Gan B. (2023). SLC7A11 expression level dictates differential responses to oxidative stress in cancer cells. Nature Communications, 14, 3673.

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Epigenetic Regulation in Breast Cells

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Human BC cells (MDA-MB-231, Hs578T, T47D, SKBR3, BT474, BT549, and MCF-7) and human breast epithelial cells (MCF‐10A) were obtained from ATCC (Manassas, Virginia, USA). All cells were cultured in DMEM (Gibco, Carlsbad, CA, USA) mixed with 10% FBS (Gibco) in with 5% CO₂ at 37°C. Cells were treated with 20 µM CpG methyltransferase (M.SssI) (Thermo Fisher Scientific, Waltham, MA, USA) or 40 µM M.SssI for 24 h.

Hu Y., He Y., Luo N., Li X., Guo L, & Zhang K. (2023). A feedback loop between lncRNA MALAT1 and DNMT1 promotes triple-negative breast cancer stemness and tumorigenesis. Cancer Biology & Therapy, 24(1), 2235768.

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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).

Stanford E.A., Wang Z., Novikov O., Mulas F., Landesman-Bollag E., Monti S., Smith B.W., Seldin D.C., Murphy G.J, & Sherr D.H. (2016). The role of the aryl hydrocarbon receptor in the development of cells with the molecular and functional characteristics of cancer stem-like cells. BMC Biology, 14, 20.

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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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