Human breast carcinoma cells (luminal types: MCF7, T47D, and ZR75; basal types: MDA-MB-231 and HS578T) were obtained from the Bioresource Collection and Research Center (BCRC; https://www.bcrc.firdi.org.tw/12092013 (accessed on 30 May 2022)). Cells were maintained in DMEM (Gibco) with 5% CO2 at 37 °C in a humidified incubator. All cell culture media were supplemented with 10% FBS (Biological Industries) and 1% PSA (penicillin G/streptomycin/amphotericin B; Biological Industries). Kinase inhibitor wortmannin (20 μM; Sigma) and kinase activator SC79 (5 μM; Sigma) were used to investigate the Akt signaling pathway. Three chemotherapy drugs, fluorouracil (5FU) (Sigma), paclitaxel (Sigma), and doxorubicin (Sigma), were used to study chemoresistance in CD44-overexpressing and knockdown cells.
Hs578t
Manufactured by RIKEN BioResource Center
Sourced in Japan
Hs578T is a human breast cancer cell line. It is derived from a primary carcinoma of the breast. The Hs578T cell line is commonly used in cancer research, specifically for studies related to breast cancer.
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Lab products found in correlation
Mcf 7, by RIKEN BioResource Center (6 mentions)
Mda mb 231, by RIKEN BioResource Center (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Mda mb 453, by RIKEN BioResource Center (2 mentions)
H184b5f5 m10, by RIKEN BioResource Center (2 mentions)
Hs578t, by Thermo Fisher Scientific (2 mentions)
Mda mb 231, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Doxorubicin, by Merck Group (1 mentions)
Fluorouracil 5 fu, by Merck Group (1 mentions)
Wortmannin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Paclitaxel, by Merck Group (1 mentions)
Plc prf 5, by RIKEN BioResource Center (1 mentions)
Hep3b, by RIKEN BioResource Center (1 mentions)
Zr 75 1, by RIKEN BioResource Center (1 mentions)
Nci h520, by RIKEN BioResource Center (1 mentions)
Hyclone, by Thermo Fisher Scientific (1 mentions)
Flow cytometry, by BD (1 mentions)
Skbr3, by RIKEN BioResource Center (1 mentions)
9 protocols using hs578t
1
Breast Cancer Cell Line Characterization
2
Comprehensive Cell Line Panel for Drug Evaluation
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MDA-MB-231, MDA-MB-468, K562, HeLa, MCF7, HCC1954, A549, COLO205, U2OS, Huh-7, U937, HepG2, KG-1, PC3, BT474, MV4-11, RS4;11, MOLM-13, WI-38, HUVEC, RPTEC, and HAoSMC were from Development Center for Biotechnology, New Taipei City, Taiwan; MDA-MB-453, T47D, ZR-75-1, ZR-75-30, MDA-MB-361, Hs578T, NCI-H520, Hep3B, PLC/PRF/5 were from Bioresource Collection and Research Center, Hsinchu, Taiwan. Cell lines were maintained in complete 10% fetal bovine serum (Biowest, Miami, FL, USA or Hyclone, Thermo Scientific, Rockford, IL, USA) and physiologic glucose (1 g/L) in DME (Sigma, St. Louis, MO, USA). Studies conducted using cell lines RPMI8226, MOLT-4, and N87; drug-resistant cell lines MES-SA/Dx5, NCI/ADR-RES, and K562R were from and tested by Xenobiotic Laboratories, Plainsboro, NJ, USA.
3
Selective Antibody Binding on HER2 Cells
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The HER2-positive cells, MDA-MB-231, MCF7 and SKBR3, and the HER2-negative Hs578T cell lines were obtained from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan) and maintained according to the manufacturer’s instructions. These cell lines (3 × 105 cells) were incubated with Herceptin for 30 min followed by incubation for 30 min with fluorescein-conjugated rabbit anti-human IgG (Acris Antibodies GmbH, Herford, Germany; 1: 10,000). The fluorescence was assayed via flow cytometry (Becton–Dickinson, San Jose, CA).
LPPC was first labeled with 3 mM fluorescent lipophilic dye DiO (Sigma-Aldrich, St. Louis, MO; 10 μl in 1 mg LPPC at a final volume of 110 μl) for 30 min and subsequently washed and resuspended as described above. Next, DiO-incorporated LPPC (20 μg) was complexed with either 2 μg of Herceptin or 2 μg of Rituximab (anti-human CD20 antibody) and then blocked with 20 μl of PEG1500 (100 mg/ml) for an additional 30 min. Various human breast tumor cells (3 × 105 cells) were incubated with 20 μg of DiO/LPPC/Herceptin or DiO/LPPC/Rituximab at 4 °C for 30 min in the dark. After the cells were washed and resuspended in 1 ml DMEM, the cells were analyzed by a flow cytometry.
LPPC was first labeled with 3 mM fluorescent lipophilic dye DiO (Sigma-Aldrich, St. Louis, MO; 10 μl in 1 mg LPPC at a final volume of 110 μl) for 30 min and subsequently washed and resuspended as described above. Next, DiO-incorporated LPPC (20 μg) was complexed with either 2 μg of Herceptin or 2 μg of Rituximab (anti-human CD20 antibody) and then blocked with 20 μl of PEG1500 (100 mg/ml) for an additional 30 min. Various human breast tumor cells (3 × 105 cells) were incubated with 20 μg of DiO/LPPC/Herceptin or DiO/LPPC/Rituximab at 4 °C for 30 min in the dark. After the cells were washed and resuspended in 1 ml DMEM, the cells were analyzed by a flow cytometry.
4
Breast Cancer Cell Culture Protocols
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Human breast cancer MCF7 cells were cultured as described37 (link). T47D and Hs578T cells were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan) and cultured in RPMI 1640 medium or DME medium (Sigma-Aldrich) supplemented with 10 μg/ml insulin (Thermo Fisher Scientific). All cells were maintained in medium supplemented with 10% fetal bovine serum (Thermo Fisher Scientific). Plasmid or siRNA transfection for all cells was performed using Lipofectamine 2000 as described37 (link). To induce EGFR signaling, EGF (Peprotech) was added to serum-free T47D culture medium with subsequent incubation for 24 h. Afatinib, AZD9291, PD153035, and AEB071 were purchased from Selleckchem, and the MEK inhibitor U0126 was obtained as described38 (link).
5
Breast Cancer Cell Culture Conditions
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The breast cancer cell lines MDA-MB-453, YMB-1, MCF-7, BT549, and Hs578T were obtained from the RIKEN BioResource Center (RIKEN BRC, Tsukuba, Japan) and Health Science Research Resource Bank (HSRRB, Osaka, Japan). MDA-MB-231 and MDA-MB-468 cells were supplied by Dr. Nishiguchi (Kyoto Pharmaceutical University, Kyoto, Japan). Cells were cultured in Leibovitz's L-15, RPMI 1640, or D-MEM medium (Wako Pure Chemical Industries, Osaka, Japan) supplemented with 10% fetal bovine serum (FBS), 100 units/mL of penicillin/streptomycin, and 2 mM glutamine. Cells were maintained at 37°C in an atmosphere of 5% CO2 and 95% air. Commercial FBS contains approximately 0.3 nM testosterone, and the final concentration of testosterone in culture medium is approximately 0.03 nM. Under high testosterone conditions, exogenous DHT (Tokyo Chemical Industry, Tokyo, Japan) was added to the medium at a final concentration of 10 nM. Under testosterone deprivation conditions, charcoal-stripped FBS (Sigma, St. Louis, MO, USA) was added to the medium instead of normal FBS, without the supplementation of 10 nM DHT.
6
Cell Culture Conditions for Cancer Research
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MDA-MB-231, Hs578T, and HCT116 were obtained from the Bioresource Collection and Research Center (BCRC, Taiwan). H1299 and 293T cells were obtained from the ATCC. Cells were grown in Dulbecco's modified Eagle's medium (DMEM) containing 10% FBS. All cells were cultured at 37 °C with 5% CO2. Cell lines were routinely tested to exclude mycoplasma contamination.
7
Culturing Human Breast Cancer Cell Lines
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Human breast cancer cell lines Hs578T, MDA-MB-231, MCF-7, and BT-474 and the human mammary epithelial cell line H184B5F5/M10 were purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). These cell lines were cultured in Dulbecco’s Modified Eagle Medium (Sigma, St. Louis, MO, USA) with 0.1 mM sodium pyruvate, 10% fetal bovine serum (Gibco, BRL, Grand Island, NY, USA), 2 mM l -glutamine, 100 U mL−1 penicillin, and 100 µg mL−1 streptomycin (Sigma-Aldrich, Saint Louis, MO, USA). The cells were maintained at 37 °C in a humidified incubator with 5% CO2.
8
Breast Cancer Cell Line Cultures
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Human breast cancer cell lines MCF-7, T-47D, Hs578T, MDA-MB-231, and nontumorigenic human breast epithelial cell line H184B5F5/M10 were originally obtained from the Bioresource Collection and Research Center. Hs578T and MDA-MB-231 were maintained in Dulbecco's Modified Eagle medium containing 10% fetal bovine serum, 1μg/ml penicillin and 1μg/ml streptomycin (Invitrogen) at 37°C in a 5% CO2 atmosphere. MCF-7 and H184B5F5/M10 were maintained in MEM-α, and T-47D was maintained in Roswell Park Memorial Institute medium 1640 with the same supplements and culture condition. And two specific inhibitors of MEK/ERK pathway, PD98059 (50μM) and U0126 (20μM, Sigma-Aldrich Co), were used as parallel experiments for 72 h as the comparison of the effect of miR-550a-3p in both MDA-MB-231 and MCF-7 cells.
9
Culturing Breast Cancer Cell Lines
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MDA-MB-231 (ATCC Cat# CRL-12532), HCC-1395 (ATCC Cat# CRL-2324), Hs578T (ATCC Cat# CRL-7849), and MCF-7 (ATCC Cat# HTB-22) cells were purchased from Bioresource Collection and Research Center (BCRC), Taiwan. MDA-MB-231, Hs578T, and MCF-7 cells were maintained in DMEM (Invitrogen, CA, USA) while HCC-1395 cells were maintained in RPMI (Invitrogen, CA, USA) containing 10% fetal bovine serum (FBS; HyClone, UT, USA) and 1× penicillin-streptomycin (Invitrogen, CA, USA) at 37°C in an atmosphere containing 5% CO2. Non-tumorigenic mammary epithelial line MCF-10A (ATCC Cat# CRL-10317) was cultured in DMEM/F-12 medium (Invitrogen, CA, USA) supplemented with 5% of horse serum (Invitrogen, CA, USA), 20 ng/mL of EGF (Pro-Spec, Rehovot, Israel), 10 μg/mL of insulin (Sigma-Aldrich, MI, USA), 0.5 μg/mL of hydrocortisone (Calbiochem, Darmstadt, Germany), 100 ng/mL of cholera toxin (Sigma-Aldrich, MI, USA), and 1% of 1× penicillin-streptomycin (Invitrogen, CA, USA) at 37°C in an atmosphere containing 5% CO2.
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