In the present study, cell lines representative of cancer types were selected as described previously (16 (link)): MCF7 and T47D for luminal A type; BT474 for luminal B type; and MDA-MB468, MDA-MB231, HCC70 and HCC1143 for TNBC. The human breast cancer cell lines were obtained from the Riken Cell Bank (MCF7; Riken BioResource Research Center, Tsukuba, Japan) and from the American Type Culture Collection (T47D, BT474, MDA-MB468, MDA-MB231, HCC70 and HCC1143; Manassas, VA, USA). Cells were cultured in RPMI-1640 medium (Wako Pure Chemical Industries, Ltd., Osaka, Japan) supplemented with 100 U/ml of penicillin, 100 U/ml of streptomycin and 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) in a humidified atmosphere containing 5% CO2 at 37°C.
Mcf 7
Manufactured by RIKEN BioResource Center
Sourced in Japan
MCF-7 is a cell line derived from a human breast adenocarcinoma. It is a widely used model system for studying breast cancer biology and evaluating the effects of various compounds on cancer cells.
Automatically generated - may contain errors
Lab products found in correlation
Mda mb 231, by RIKEN BioResource Center (28 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (7 mentions)
Hs578t, by RIKEN BioResource Center (6 mentions)
Mda mb 231, by Thermo Fisher Scientific (5 mentions)
Mcf 7, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Sartorius (4 mentions)
Mda mb 453, by RIKEN BioResource Center (4 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Bt 474, by RIKEN BioResource Center (4 mentions)
H184b5f5 m10, by RIKEN BioResource Center (3 mentions)
Skbr3, by RIKEN BioResource Center (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Hs578t, by Thermo Fisher Scientific (3 mentions)
Mda mb 231, by American Type Culture Collection (2 mentions)
Mda mb 468, by American Type Culture Collection (2 mentions)
Caco 2, by RIKEN BioResource Center (2 mentions)
Nih3t3, by RIKEN BioResource Center (2 mentions)
Paclitaxel, by Merck Group (2 mentions)
54 protocols using mcf 7
1
Breast Cancer Cell Line Culture Protocol
2
Cancer Cell Line Cultivation and Maintenance
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The human cancer cell lines HeLaS3, KBvin, MCF-7, HepG2, NCI-H460 and NCI-H460/MX20 were grown in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) and 1/100 (v/v) penicillin/streptomycin at 37 °C under 5% carbon dioxide. The medium was routinely changed every two days. As cell confluence reached 80% in the medium, the cells were treated with 0.25% of trypsin-EDTA for the next passage. Human cervical carcinoma cell line HeLaS3, human non-small-cell lung carcinoma cell line NCI-H460, human liver carcinoma cell line HepG2 and human breast cancer cell line MCF7 were purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan). The multidrug-resistant human cervical cancer cell line KB-vin was kindly provided by Dr. Kuo-Hsiung Lee (University of North Carolina, Chapel Hill, NC, USA) and maintained with 100 nM of vincristine each week. MDR human non-small-cell lung carcinoma cell line NCI-H460/MX20 was the resistant cell line selected from NCI-H460. It was treated with gradually increasing concentrations of mitoxantrone and maintained in 20 nM of mitoxantrone.
Note: all tested compounds were dissolved in DMSO and the final concentrations of DMSO in all experiments were less than 0.1%.
Note: all tested compounds were dissolved in DMSO and the final concentrations of DMSO in all experiments were less than 0.1%.
3
Isolation and Characterization of Extracellular Vesicles from MCF-7 Cells
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MCF-7 (Bioresource Collection and Research Center, Hsinchu, Taiwan) cells were cultured and grown in 15 cm dishes (Corning). When cells reached approximately 80% confluence, cells were washed with phosphate buffered saline (PBS) (Gibco) and transferred to fresh media containing 1% EV-free serum (Gibco) for 24 h. Media was then collected and subjected to centrifugation at 500×g for 5 min to pellet cells, then 2000×g for 15 min to remove cellular debris (Eppendorf, Hamburg, Germany). The supernatant was then transferred to ultracentrifuge tubes (Beckman Coulter, Pasadena, CA, USA) and ultracentrifuged (Beckman Coulter) at 10,000×g for 30 min twice to pellet and remove large vesicles. The supernatant was then transferred to new ultracentrifuge tubes and ultracentrifuged twice at 100,000×g for 60 min to pellet EVs. EVs were resuspended in PBS and then stored at 4 °C. Protein content of EVs was determined by BCA (Thermo Fisher Scientific) and treatment dosing was determined by EV protein concentration. Particle size was measured using qNano Gold (Izon, Science Ltd., Burnside, New Zealand) by Tunable Resistive Pulse Sensing. Data was recorded and analyzed using Izon Control Suite Software (version 2.2.2.111). Particle size reported are representative of three separate measurements of EVs.
4
MCF-7 Cell Hypoxia-Reoxygenation Protocol
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Human breast adenocarcinoma cell line MCF-7 was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan). MCF-7 cells were maintained in Dulbecco's modified Eagle's medium (DMEM, Life Technologies, Carlsbad, CA, USA) supplemented with 10% (v/v) fetal bovine serum (Life Technologies) and 1% antibiotic solution (penicillin-streptomycin-amphotericin solution, Biological Industries, Beit-Haemek, Israel) at 37°C in a humidified incubator under 5% CO2. For hypoxia cultures, cells were incubated in a hypoxia chamber (InVivO2-200, Ruskinn Technology, Leeds, UK) for 24 h with 0.5% O2, 5% CO2 and residual N2. After 24 h of hypoxia, cells were shifted to a humidified incubator with 5% CO2 and 95% air, and incubated at 37°C. The samples were harvested at selected time points after reoxygenation.
5
Culturing Human Breast Cancer Cell Lines
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Human breast cancer cell lines (BCCs), MCF-7 and MDA-MB-231, were purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan) and cultured in Dulbecco’s modified Eagle medium (DMEM) and Roswell Park Memorial Institute Medium-1640 (RPMI-1640), respectively, and supplemented with 10% fetal bovine serum (FBS; Gibco BRL, Carlsbad, CA). Both cell lines were maintained at 37°C in a humidified 5% CO2 incubator.
6
Stimuli-Responsive Polymeric Nanoparticles for Cancer Therapy
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Acrylic acid (AA) and N-isopropylacrylamide (NIPAAm) were supplied from Acros Organics (Geel, Flemish Region, Belgium). Both 2-(Dodecylthiocarbonothioylthio)-2-methylpropionic acid (DMP) as a chain transfer agent (CTA) and 2,2′-azobisisobutyronitrile (AIBN) initiator were obtained from Aldrich (St. Louis, MO, USA). An anticancer drug, camptothecin (CPT), was also obtained from Acros Organics. Dimethyl sulfoxide (DMSO) was obtained from J.T. Baker (Philadelphia, PA, USA). Chitosan (CS) (C&B Co., Taipei, Taiwan) was purified according to the previous procedure [66 (link)]. The purified CS had an average molecular weight of 24.5 kDa and a degree of deacetylation (DDA, the fraction of amino group on C-2 position) of 0.94. The mouse fibroblast cells (NIH 3T3), human intestinal epithelial cells (Caco-2), non-small human lung carcinoma cells (H460), and human breast cancer cells (MCF-7) were provided by the Bioresource Collection and Research Center, Hsinchu, Taiwan.
7
Plasmid Transfection of A549 Cells
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Cell lines, MCF-7 and A549, were purchased from the Bioresource Collection and Research Center (BCRC). Standard culture procedures and conditions were followed, according to instructions from the BCRC. For plasmid transfection, 1 × 106 A549 cells were seeded on a 10 cm2 plate. After overnight culture, 16 µg pmWasabi-ANT2 plasmid DNA was transfected using LipofectAMINETM3000 (ThermoFisher Scientific, Carlsbad, CA, USA), according to the recommended protocol from the manufacturer. Cells were cultured for 48 h and harvested for antibody pulldown or immunocytochemical studies.
8
Evaluation of Breast Cancer Cell Lines
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Human breast cancer cell (MCF-7) and human breast epithelial cell (H184B5F5/M10) were obtained from Bioresource Collection and Research Center, Taiwan Food Industry Research and Development Institute (Hsinchu, Taiwan). Minimum Essential Medium (MEM) medium, α-MEM medium, fetal bovine serum (FBS), HEPES solution, sodium bicarbonate solution, sodium pyruvate, phosphate buffered saline (PBS), Hank’s balanced salt solution (HBSS) and 0.25% trypsin-EDTA were all from HyClone Co. (Logan, UT, USA).
Specific pathogen free (SPF) male BALB/c nude mice (four-weeks old) were purchased from the National Laboratory Animal Center (Taipei, Taiwan). The diet (LabDiet) was from LabDiet Co. (St. Louis, MO, USA). Both mouse EGF and VEGF ELISA kits were from Koma Biotech Co. (Seoul, Korea). Paclitaxel was from Sigma-Aldrich Co., while estradiol cypionate was from Da-Fong Drug Co. (Chanhua, Taiwan). The use of animals (mice) and the associated experimental procedures were approved by Fu Jen University Animal Care and Use Committee (Ethical permission approval number and date: A10704 and 23 April 2018).
Specific pathogen free (SPF) male BALB/c nude mice (four-weeks old) were purchased from the National Laboratory Animal Center (Taipei, Taiwan). The diet (LabDiet) was from LabDiet Co. (St. Louis, MO, USA). Both mouse EGF and VEGF ELISA kits were from Koma Biotech Co. (Seoul, Korea). Paclitaxel was from Sigma-Aldrich Co., while estradiol cypionate was from Da-Fong Drug Co. (Chanhua, Taiwan). The use of animals (mice) and the associated experimental procedures were approved by Fu Jen University Animal Care and Use Committee (Ethical permission approval number and date: A10704 and 23 April 2018).
9
Culturing ER-positive Breast Cancer Cell Lines
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Human breast cancer cell lines, ER-positive luminal A (MCF-7 and T-47D), were bought from the Bioresource Collection and Research Center, Taiwan. MCF-7 was cultured in RPMI1640 medium (Gibco BRL, Grand Island, NY, USA) supplemented with 8% fetal bovine serum (FBS). T-47D was maintained in RPMI1640 medium using 8% FBS, containing 1.5-g/L sodium bicarbonate, 4.5-g/L glucose, 10-mM HEPES, and 1.0-mM sodium pyruvate and 0.2 I.U. bovine insulin per ml. The cell lines were kept in a humidified atmosphere with 5% CO2 at 37˚C.
10
Stereotactic Breast Biopsy for Microcalcifications
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Women with Breast Imaging Reporting and Data System (BI-RADS) category 4 or 5 due to suspicious malignant MC on screening or diagnostic mammography were recommended for stereotactic breast biopsy. Biopsies of breast MC and adjacent normal tissue (non-MC) were obtained by the stereotactic 10-gauge vacuum-assisted breast biopsy system (Vacora, Bard Medical Systems, Tempe, AZ). In general, 15 to 20 specimens per lesion were obtained. Women were included if mammography between January 2010 and December 2011 showed that MC specimens were insufficient for clinical pathology diagnosis.
Breast cancer cell lines, MCF-7 (BCRC#60436) and MDA-MB-231 (BCRC#60549) from the Bioresource Collection and Research Center (BRCR, Hsinchu, Taiwan) were cultured in DMEM supplemented with 10% fetal bovine serum (Invitrogen) in a humidified atmosphere of 95% air and 5% CO2 at 37°C.
Breast cancer cell lines, MCF-7 (BCRC#60436) and MDA-MB-231 (BCRC#60549) from the Bioresource Collection and Research Center (BRCR, Hsinchu, Taiwan) were cultured in DMEM supplemented with 10% fetal bovine serum (Invitrogen) in a humidified atmosphere of 95% air and 5% CO2 at 37°C.
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