ES2 (CRL-1978) and OVCAR3 (HTB-161) OC cells were obtained from the American Type Culture Collection (ATCC) (Manassas, VA). Cells were maintained in DMEM supplemented with 10% fetal calf serum, penicillin, streptomycin, amphotericin, L-glutamine, sodium pyruvate, vitamins and non-essential amino acids (Biological Industries, Beit Haemek, Israel). Cell cultures were incubated at 37°C in humidified atmosphere of 95% air and 5% CO2.
Ovcar3 htb 161
Manufactured by American Type Culture Collection
Sourced in United Kingdom
OVCAR3 (HTB-161) is a human ovarian adenocarcinoma cell line derived from the malignant ascites of a patient with progressive adenocarcinoma of the ovary. This cell line is commonly used in cancer research and drug discovery studies.
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Lab products found in correlation
Caov 3 htb 75, by American Type Culture Collection (2 mentions)
P06 07300, by PAN Biotech (2 mentions)
Paclitaxel, by Merck Group (1 mentions)
A2780, by European Collection of Cell Cultures (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Gemini Bio (1 mentions)
Mda mb 231 htb 26, by American Type Culture Collection (1 mentions)
Mda mb 231, by Corning (1 mentions)
Ovcar 3, by Corning (1 mentions)
Accutase cell detachment solution, by Merck Group (1 mentions)
Fb 1550, by Biosera (1 mentions)
A5955, by Merck Group (1 mentions)
Skov3 htb 77, by American Type Culture Collection (1 mentions)
Bovine insulin, by Merck Group (1 mentions)
Pcr mycoplasma detection kit, by Applied Biological Materials (1 mentions)
Rpmi 1640 medium, by Euroclone (1 mentions)
Fetal bovine serum (fbs), by GE Healthcare (1 mentions)
Caov 3, by Euroclone (1 mentions)
Dulbecco modified eagle medium (dmem), by Euroclone (1 mentions)
9 protocols using ovcar3 htb 161
1
Cultivation of ES2 and OVCAR3 OC Cells
2
Ovarian Cancer Cell Lines Protocol
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The human ovarian carcinoma cell line A2780 was obtained from the European Collection of Cell Cultures. The human ovarian adenocarcinoma cell lines OVCAR‐3 (HTB‐161) and Caov‐3 (HTB‐75) were obtained from the American Type Culture Collection (ATCC). Spontaneously transformed mouse ovarian surface epithelium (MOSE) were developed from C57BL/6 mice and characterized previously.17 MOSE cells were transduced with firefly luciferase lentiviral particles (GeneCopoeia) (MOSE‐LTICv) to facilitate live in vivo imaging of tumor outgrowth. Paclitaxel (Sigma Aldrich, Rehovot, Israel) stock solution was prepared in DMSO and diluted in cell culture media immediately prior to use so that final DMSO concentration did not exceed 0.1%.
3
Culturing Cell Lines for Cancer Research
4
Characterization of HGSOC Cell Lines
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The PEO1 (#10032308) and PEO4 (#10032309) cell lines were purchased from the European Collection of Authenticated Cell Cultures, England. The OVCAR3 (#HTB-161) cell line was purchased from American Type Culture Collection, England. PEO1, PEO4 and OVCAR3 cell lines have previously been phenotypically and genotypically characterised as representative of HGSOC [36 (link),37 ]. For routine culture, PEO1 and PEO4 cells were maintained in RPMI 1640 growth medium (#31870, Thermo Fisher Scientific, UK), supplemented with 10% foetal bovine serum (FBS) (#FB-1550, Biosera, USA), 2 mM glutamine (#25030081, Thermo Fisher Scientific, UK), 1% antibacterial antimycotic solution (ABAM) (#A5955, Merck, UK). For routine culture, OVCAR3 cells were maintained in RPMI 1640 growth medium supplemented with 20% FBS (#FB-1550, Biosera, USA), 2 mM glutamine, 0.1% insulin (#I0516, Merck) and 1% ABAM (#A5955, Merck). All cells were cultured in TC treated Corning® T-25 (#CLS430639, Merck), T-75 (#CLS430641U, Merck), or T-175 (#CLS431080, Merck) cell culture flasks and incubated at 37 °C in a humidified 5% CO2 chamber. Growth medium was replaced every 2–3 days and cells were sub-cultured, using Accutase® cell detachment solution (#SCR005, Sigma–Aldrich), to ensure 70–80% confluency.
5
Cell Line Acquisition for Ovarian Cancer
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IGROV1 and OVMANA cells were obtained from the National Cancer Institute (NCI) and Japanese Collection of Research Bioresources, respectively. SKOV3 (HTB-77) and OVCAR3 (HTB-161) cells were purchased from American Type Culture Collection.
6
Cultivation of Ovarian Cancer Cell Lines
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Ovarian cancer cell lines PEO1 (ECACC 10032308), PEO4 (ECACC 10032309), PEO14 (ECACC 10032311) were purchased from the American European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK), whereas ovarian cancer cell lines OVCAR-3 (HTB-161) and Caov-3 (HTB-75) were ordered from American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were cultivated according to manufacturers’ instructions at 37 °C and in the presence of 5% CO2. ATCC formulated RPMI 1640 medium (Euroclone, Milan, Italy), supplemented with 10% FBS (GE Healthcare, Chicago, IL) was used for maintenance of PEO1, PEO4, and PEO14 cell lines. RPMI 1640 medium supplemented with 20% FBS and 0.01 mg/ml bovine insulin (Merck, Darmstadt, Germany) was used for cultivation of OVCAR-3 cells. Caov-3 cells were grown in DMEM (Euroclone) supplemented with 10% FBS. For all cell lines, 100 U/ml penicillin (Lonza, Basel, Switzerland), 100 mg/mL streptomycin (Lonza), and 250 ng/mL Amphotericin-B (Merck) were added to the culture medium. Mycoplasma contamination was routinely tested using a mycoplasma PCR detection kit (ABM, Richmond, BC, Canada).
7
Breast Cancer Cell Line Culturing
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Cell lines from OCCC (ES2; CRL-1978) and OSC (OVCAR-3; HTB-161) were obtained from American Type Culture Collection (ATCC). Cells were maintained at 37 °C in a humidified 5% CO2 atmosphere. Cells were cultured in DMEM (41965–039, Gibco, Life Technologies) containing 4.5 g/L of D-glucose and 0.58 g/L of L-glutamine supplemented with 1% fetal bovine serum (FBS S 0615, Merck), 1% antibiotic-antimycotic (AA) (P06–07300, PAN Biotech).
Prior to any experiment, cells were synchronized under starvation (culture medium without FBS) for 8 h at 37 °C and 5% CO2.
After 24 h of conditions exposure, the medium was changed and fresh conditions were added, with the exception of proliferation curve and cell cycle analysis in which the medium was not changed.
Prior to any experiment, cells were synchronized under starvation (culture medium without FBS) for 8 h at 37 °C and 5% CO2.
After 24 h of conditions exposure, the medium was changed and fresh conditions were added, with the exception of proliferation curve and cell cycle analysis in which the medium was not changed.
8
Culturing of Ovarian and Breast Cancer Cells
9
Hypoxia and L-Cysteine Effects on Cell Lines
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Cell lines ES2 (CRL-1978), OVCAR-3 (HTB-161) and OVCAR-8 (CVCL-1629) were obtained from American Type Culture Collection (ATCC). Cell line A2780 sensitive (93112519) and A2780 cisplatin resistant (93112517) were obtained from Sigma Aldrich. Cells were maintained at 37 °C in a humidified 5% CO2 atmosphere. ES2, OVCAR3 and OVCAR8 cell lines were cultured in DMEM (41965–039, Gibco, Life Technologies) supplemented with 1% FBS (S 0615, Merck), 1% antibiotic-antimycotic (AA) (P06–07300, PAN Biotech). A2780 parental and A2780 cisR cells were cultured in RPMI 1640 (BE12-167F, Lonza) supplemented with 0.58 g/L of L-glutamine, 1% FBS (S 0615, Merck) and 1% antibiotic-antimycotic (AA) (P06-07300, PAN Biotech).
Cells were exposed to 0.4 mM L-Cysteine (102839, Merck) and/or exposed to hypoxia-induced conditions with 0.1 mM cobalt chloride (C8661, Sigma-Aldrich). Cobalt is a hypoxia mimicking agent used in in vivo53 (link) and in vitro54 –56 (link) studies. Chemically, CoCl2 reacts with oxygen impairing its dissolution and oxygenation of aqueous solutions57 and is a way of impairing the availability of oxygen in culture media.
Prior to any experiment, cells were synchronized under starvation (culture medium without FBS) for 8 h at 37 °C and 5% CO2.
Cells were exposed to 0.4 mM L-Cysteine (102839, Merck) and/or exposed to hypoxia-induced conditions with 0.1 mM cobalt chloride (C8661, Sigma-Aldrich). Cobalt is a hypoxia mimicking agent used in in vivo53 (link) and in vitro54 –56 (link) studies. Chemically, CoCl2 reacts with oxygen impairing its dissolution and oxygenation of aqueous solutions57 and is a way of impairing the availability of oxygen in culture media.
Prior to any experiment, cells were synchronized under starvation (culture medium without FBS) for 8 h at 37 °C and 5% CO2.
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