The ovarian cancer cell lines (OCCLs) A2780, IGROV-1, OVCAR-8, and SK-OV-3 were used in this work. OVCAR-8 and SK-OV-3 were acquired from the American Type Culture Collection (ATCC), A2780 from the European Collection of Authenticated Cell Cultures (ECACC), and IGROV-1 from Merck Millipore, Burlington, MA, USA. IGROV-1 and A2780 were cultured in RPMI 1640 medium (Gibco, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gibco) and 1% penicillin/streptomycin (Gibco). SK-OV-3 and OVCAR-8 were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco) supplemented with 10% FBS and 1% penicillin/streptomycin. All cells were incubated at 37 °C in a 5% CO2 atmosphere. The presence of mycoplasma was routinely checked with the MycoAlert kit (Lonza, Basel, Switzerland), and only mycoplasma-free cells were used in the experiments.
A2780
Manufactured by European Collection of Authenticated Cell Cultures
Sourced in United Kingdom, United States, Japan
The A2780 is a laboratory cell line derived from a human ovarian carcinoma. It is a commonly used in vitro model for the study of ovarian cancer.
Automatically generated - may contain errors
Lab products found in correlation
Skov3, by American Type Culture Collection (16 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (12 mentions)
A2780, by Thermo Fisher Scientific (9 mentions)
Ovcar3, by American Type Culture Collection (8 mentions)
A2780cis, by European Collection of Authenticated Cell Cultures (7 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Skov3, by European Collection of Authenticated Cell Cultures (5 mentions)
Skov3, by Thermo Fisher Scientific (5 mentions)
Caov3, by American Type Culture Collection (5 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (5 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Mcf 7, by European Collection of Authenticated Cell Cultures (4 mentions)
Mda mb 231, by European Collection of Authenticated Cell Cultures (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (3 mentions)
Cisplatin, by Merck Group (3 mentions)
Igrov 1, by Thermo Fisher Scientific (2 mentions)
Igrov 1, by Merck Group (2 mentions)
Ovcar8, by Thermo Fisher Scientific (2 mentions)
Mycoalert kit, by Lonza (2 mentions)
39 protocols using a2780
1
Ovarian Cancer Cell Line Characterization
2
Cisplatin-Resistant Ovarian Cancer Cell Lines
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The human ovarian cancer cell lines, A2780 and SKOV3, were obtained from the European Collection of Authenticated Cell Cultures (93112519, ECACC) and the American Type Culture Collection (HTB-77, ATCC), respectively and maintained in RPMI-1640 (10–040-CVR, Corning, Inc.) and McCoy 5A medium (16600-108, Gibco; Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS) at 37°C in 5% CO2. The cisplatin-resistant cells, SKOV3/DDP and A2780/DDP, were established by exposing the cells to gradually increasing concentrations (from 0 to 25 µg/ml) of cisplatin for 6 months (data not shown). Cisplatin was purchased from Hansoh Pharmaceutical Co., Ltd. 3-Methyladenine (3-MA), rapamycin (Rapa), compound C and monodansylcadaverine (MDC) were purchased from Sigma-Aldrich. The primary antibody to TRP14 (ab121725/dilution, 1:500) was purchased from Abcam. The primary antibodies for ATG5 (#12994/dilution, 1:1,000), Beclin1 (#3495/dilution, 1:1,000), LC3B (#3868/dilution, 1:1,000), p-5′ AMP-activated protein kinase (AMPK; #2537/dilution, 1:1,000), p-mammalian target of rapamycin (mTOR; #5536/dilution, 1:1,000), p-p70S6K (#9204/dilution, 1:1,000), AMPK (#5832/dilution, 1:1,000), mTOR (#2983/dilution, 1:1,000) and p70S6K (#2708 /dilution, 1:1,000) were obtained from Cell Signaling Technology Inc. The antibody for GAPDH (sc-47724) was purchased from Santa Cruz Biotechnology.
3
Cell Line Culture Protocols
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A2780 (#93112519) and A2780Cis (#93112517) cell lines were purchased from European Collection of Authenticated Cell Cultures (ECACC, Sigma-Aldrich, St. Louis, MO, USA). VH10, U2OS, SKOV-3, and HEK293T cell lines were purchased from American Type Culture Collection (ATCC®, Manassas, VA, USA). The PEO1 and PEO1.C2-4 were a gift from Dr. Kumar Sanjiv (cell line identity had been confirmed by Sanger sequencing), the BJhTERT cells were provided by W. Hahn (Dana–Farber Cancer Institute) and OVCAR-4 cells were a kind gift from Dr. Kaisa Lehti. A2780, A2780cis, PEO1, PEO1.C2-4, and OVCAR-4 were cultured in RPMI 1640 with GlutaMAX (#61870 Thermo Fisher Scientific, Waltham, MA, USA) with supplementation of 2% pyruvate (#11360070 Thermo Fisher, for PEO1 and PEO1.C2-4). U2OS were cultured in DMEM with GlutaMAX (#31966-021, Thermo Fisher Scientific), HEK293T cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM; #12440, Thermo Fisher Scientific) and SKOV-3 cells were cultured in McCoy 5A (#16600082, Thermo Fisher Scientific). All media was supplemented with 10% fetal bovine serum (FBS; #10500, Thermo Fisher Scientific), except for U2OS with 5% FBS, and 100 U/mL penicillin– streptomycin antibiotics (#15070, Thermo Fisher Scientific). Cell were grown at 37 °C in 5% CO2 humidified incubators.
4
Cell Lines and Culturing Conditions
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The cancer cell lines utilized for our investigations, including human ovarian carcinoma (A2780), human cervix carcinoma (Hela), human breast adenocarcinomas (MCF-7 and MDA-MB-231), and mouse embryonic fibroblast cell line (NIH/3T3) were purchased from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK). Another cervical cell line (SiHa) was acquired from the American Tissue Culture Collection (Manassas, VA, USA). For optimal growth, all cell lines were maintained in minimal essential medium (MEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 1% MEM non-essential amino acid solution (MEM-NEAA), and 1% penicillin/streptomycin mixture, at 37 °C, in a humidified incubator, containing 5 % carbon dioxide (CO2). All the utilized media and supplements were purchased from Lonza Group Ltd. (Basel, Switzerland). Unless otherwise specified, the chemicals and kits used for the experiments were purchased from Merck Life Science Ltd. (Budapest, Hungary).
5
Culturing Ovarian Cancer Cell Lines
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Human OC cell lines OVCAR-3 (adenocarcinoma), SK-OV-3 (adenocarcinoma), CAOV-3 (adenocarcinoma) and ES-2 (clear cell carcinoma) cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Human ovarian carcinoma cell line A2780 was purchased from the European Collection of Authenticated Cell Cultures (ECACC, UK). OVCAR-3 and A2780 cells were cultured in RPMI-1640 and SK-OV-3 and CAOV-3 cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM, 4.5 g/L glucose, Corning Inc., Manassas, VA, USA) supplemented with 10% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA, USA). ES-2 cells were cultured in McCoy’s 5A medium (Sigma, Saint Louis, MO, USA) supplemented with 10% FBS. Non-tumorous human ovarian surface epithelial cells (HOSEpiC) as control cells were obtained from Guangzhou Jennio Biotech Co., Ltd (Guangzhou, Guangdong, China) and cultured in RPMI-1640 medium supplemented with 10% FBS. Dulbecco's Modified Eagle's Medium (DMEM), ethylenediaminetetraacetic acid (EDTA),phenylmethylsulfonyl fluoride (PMSF)
6
Cell Line Authentication and Validation
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The Ca9-22, HEL, NUGC-3, and RPMI8226 cell lines were obtained from the Japanese Collection of Research Bioresources. The 786-O, CFPAC-1, DU145, HCC1599, HCC1806, HCC38, HCT116, HL-60, K-562, MSTO-211H, MV-4-11, NCI-H460, NCI-H2170, and THP-1 cell lines were obtained from the ATCC. The A2780, COLO 792, and DOK cell lines were obtained from the European Collection of Authenticated Cell Cultures. The A549, A673, BHL-89, and MCF-7 cell lines were obtained from DS Pharma Biomedical Co., Ltd. Short-tandem repeat-based DNA profiling was used to reauthenticate cell lines. All cell lines were tested for mycoplasma contamination.
7
Ovarian Cancer Cell Line Cultivation
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Ovarian cancer cell lines PEO1 and PEO4 cells were obtained from the American Type Culture Collection. A2780, A2780-cis and SKOV-3 cells were obtained from the European Collection of Authenticated Cell Cultures. Cells were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (SIGMA) (for A2780 and A2780-cis), RPMI medium (SIGMA,) containing Sodium Pyruvate (SIGMA) 2 mM (for PEO1 and PEO4) or McCoy’s 5A Modified medium (SIGMA) (for SKOV-3) supplemented with 10% heat-inactivated iron-supplemented donor bovine serum (Gibco, Life Technologies) and penicillin/streptomycin (SIGMA) (with 10,000 units penicillin and 10 mg streptomycin/mL) in 37 °C and 5% CO2 atmosphere. Cisplatin (1 µM) was added to A2780-cis culture media every two to three passages. Cell line authentication was conducted using the Promega Powerplex® 16-short-tandem-repeat system. All cell lines were regularly screened for mycoplasma infection using a MycoProbe® Mycoplasma Detection Kit (R&D Systems). Cells in logarithmic growth phase were used for experiments.
8
Epithelial Ovarian Cancer Cell Lines
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SKOV-3 and OVCAR-3 cells were obtained from the American Type Culture Collection. A2780 and A2780cis cells were purchased from the European Collection of Authenticated Cell Cultures and Addexbio Technologies, respectively. A2780 are defined as chemosensitive (cisplatin-sensitive) cells and as a type of EC in the epithelial ovarian cancer subtype (37 (link)). A2780cis, SKOV-3 and OVCAR-3 are chemoresistant cells and are classified as EC, CC and HGSC in the epithelial ovarian cancer subtype, respectively (37 (link),38 (link)). All cells were maintained in RPMI-1640 medium (Thermo Fisher Scientific, Inc.) with different supplements. Medium for A2780, A2780cis and SKOV-3 cells was supplemented with 10% FBS (Thermo Fisher Scientific, Inc.), 1% penicillin/streptomycin (Thermo Fisher Scientific, Inc.) and 1% L-glutamine (Thermo Fisher Scientific, Inc.). Medium for OVCAR-3 cells was supplemented with 20% FBS, 1% penicillin/streptomycin and 1% L-glutamine. For A2780cis cells, 1 µM cisplatin (Sigma-Aldrich; Merck KGaA) was added into RPMI-1640 complete medium every two passages to maintain cisplatin resistance. All cells were maintained at 37°C in a 5% CO2 humidified incubator.
9
Detailed Cell Line Sourcing and Culturing
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SKOV-3, SW626, and UWB1.289 cells were obtained from the American Type Culture Collection. OVCAR8 cells were obtained from the MDACC-characterized cell line core. A2780 cells were purchased from European Collection of Authenticated Cell Cultures. The OV2008 and C13* cell lines were gifts from Professor Benjamin K. Tsang of the Ottawa Health Research Institute (Ottawa, Canada).44 (link) iKRAS cells, primary murine pancreatic cells established from p48 Cre_tetO_LKrasG12D ROSA_rtTAL+p53L+ mice, were gifted by Professor Anirban Maitra (MDACC, Houston, TX, USA). All cell lines were tested monthly for Mycoplasma using PCR and were not passaged more than 30 times.
SKOV3 cells were cultured in McCoy’s 5A medium supplemented with 10% FBS. SW626 cells were cultured in L-15 medium supplemented with 10% fetal bovine serum (FBS). UWB1.289 cells were cultured in a medium containing RPMI-1640 and MEGM (Lonza, CC-3150) mixed at a 1:1 ratio with 3% FBS. SW626 cells were cultured in L-15 medium supplemented with 10% FBS. OVCAR8, A2780, OV2008, C13*, and iKRAS cells were cultured in RPMI-1640 medium supplemented with 10% FBS. All cell lines except SW626 were cultured at 37 °C in a 5% (v/v) CO2 atmosphere. SW626 cells were cultured at 37 °C in a free gas exchange with atmospheric air.
SKOV3 cells were cultured in McCoy’s 5A medium supplemented with 10% FBS. SW626 cells were cultured in L-15 medium supplemented with 10% fetal bovine serum (FBS). UWB1.289 cells were cultured in a medium containing RPMI-1640 and MEGM (Lonza, CC-3150) mixed at a 1:1 ratio with 3% FBS. SW626 cells were cultured in L-15 medium supplemented with 10% FBS. OVCAR8, A2780, OV2008, C13*, and iKRAS cells were cultured in RPMI-1640 medium supplemented with 10% FBS. All cell lines except SW626 were cultured at 37 °C in a 5% (v/v) CO2 atmosphere. SW626 cells were cultured at 37 °C in a free gas exchange with atmospheric air.
10
Ovarian Cancer Cell Line Cultivation
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Ovarian cancer cell lines (OCCLs) IGROV-1, OVCAR-8, SK-OV-3, and A2780 were used in this work. OVCAR-8 and SK-OV-3 were obtained from the American Type Culture Collection (ATCC), A2780 from the European Collection of Authenticated Cell Cultures (ECACC), and IGROV-1 from Merck Millipore. A2780 and IGROV-1 were cultured in RPMI 1640 medium (Gibco, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gibco, Waltham, MA, USA) and 1% penicillin/streptomycin (Gibco, Waltham, MA, USA). OVCAR-8 and SK-OV-3 were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Waltham, MA, USA) supplemented with 10% FBS and 1% penicillin/streptomycin. All cells were incubated at 37 °C in a 5% CO2 atmosphere. The presence of mycoplasma was routinely checked with the MycoAlert kit (Lonza, Basel, Switzerland) and only mycoplasma-free cells were used in the experiments.
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