The human ovarian epithelial cancer cells, A2780CP20, HEYA8, and HEYA8.MDR, were provided by Dr. Anil K. Sood (MD Anderson Cancer Center, Houston, TX, USA) and have been described elsewhere [78 (link)]. A2780 was purchased from the European Collection of Cell Cultures (ECACC, Porton Down, Salisbury, UK), and OV90 and OVCAR3 from the American Type Culture Collection (ATCC, Manassas, VA, USA)—both of which provide authenticated cell lines. OV90CIS and OVCAR3CIS cells were generated by exposing OV90 and OVCAR3 to increasing concentrations of cisplatin (CIS; Sigma-Aldrich, St. Louis, MO, USA). Cells were maintained in RPMI1640 (A2780, A2780CP20, HEYA8, and HEYA8.MDR; HyClone, GE Healthcare Life Sciences, Logan, UT, USA), RPMI1640 + insulin (0.01 mg/mL, OVCAR3 and OVCAR3CIS; Sigma) or M199/MCDB-105 (OV90 and OV90CIS; Gibco, Thermo Fisher Scientific, Grand Island, NY/Sigma) medium supplemented with 10% fetal bovine serum (FBS; HyClone) and 0.1% antibiotic/antimycotic solution (HyClone) at 37 °C in 5% CO2 and 95% air. All cell lines were screened for mycoplasma using the LookOut® Mycoplasma PCR detection kit (Sigma), and authenticated by Promega (Madison, WI, USA) and ATCC using Short Tandem Repeat (STR) analysis. In vitro assays were performed at 70%–85% cell density.
Ovcar3
Manufactured by Merck Group
Sourced in United States
OVCAR3 is a human ovarian adenocarcinoma cell line. It is a well-established in vitro model used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Ovcar3, by American Type Culture Collection (13 mentions)
Skov3, by Merck Group (7 mentions)
Skov3, by American Type Culture Collection (5 mentions)
Rpmi 1640, by Merck Group (4 mentions)
Cisplatin, by Merck Group (3 mentions)
Insulin, by Merck Group (3 mentions)
Bovine insulin, by Merck Group (3 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (3 mentions)
Streptomycin, by Merck Group (3 mentions)
Penicillin, by Merck Group (3 mentions)
L glutamine, by Merck Group (3 mentions)
Skov3, by National Collection of Authenticated Cell Cultures (3 mentions)
Ovcar3, by National Collection of Authenticated Cell Cultures (3 mentions)
Oaw42, by Merck Group (3 mentions)
A2780, by European Collection of Cell Cultures (2 mentions)
Hela cells, by Merck Group (2 mentions)
Lncap, by Merck Group (2 mentions)
Authenticated cancer cell lines, by Merck Group (2 mentions)
Ovcar3cis, by Merck Group (2 mentions)
A2780, by Thermo Fisher Scientific (2 mentions)
25 protocols using ovcar3
1
Ovarian Cancer Cell Line Establishment
2
Developing Novel Proteasome Inhibitors for Cancer Treatment
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Example 8
New proteasome inhibitors may be developed not only for treating conditions mediated by senescent cells, but also conditions mediated by cancer cells.
The ability of compounds to specifically kill cancer cells can be tested in assays using other established cell lines. These include HeLa cells, OVCAR-3, LNCaP, and any of the Authenticated Cancer Cell Lines available from Millipore Sigma, Burlington Mass., U.S.A. Compounds specifically kill cancer cells if they are lethal to the cells at a concentration that is at least 5-fold lower, and preferably 25- or 100-fold lower than a non-cancerous cell of the same tissue type. The control cell has morphologic features and cell surface markers similar to the cancer cell line being tested, but without signs of cancer.
In vivo, compounds are evaluated in flank xenograft models established from sensitive SCLC (H889) and hematologic (RS4; 11) cell lines, or using other tumor-forming cancer cell lines, according to what type of cancer is of particular interest to the user. When dosed orally or intravenously, compounds induce rapid and complete tumor responses (CR) that are durable for several weeks after the end of treatment in all animals bearing H889 (SCLC) or RS4; 11 (ALL) tumors. Similar treatment of mice bearing H146 SCLC tumors can induce rapid regressions in the animals.
3
Evaluating Novel Proteasome Inhibitors for Cancer Treatment
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Example 8
New proteasome inhibitors may be developed not only for treating conditions mediated by senescent cells, but also conditions mediated by cancer cells.
The ability of compounds to specifically kill cancer cells can be tested in assays using other established cell lines. These include HeLa cells, OVCAR-3, LNCaP, and any of the Authenticated Cancer Cell Lines available from Millipore Sigma, Burlington Mass., U.S.A. Compounds specifically kill cancer cells if they are lethal to the cells at a concentration that is at least 5-fold lower, and preferably 25- or 100-fold lower than a non-cancerous cell of the same tissue type. The control cell has morphologic features and cell surface markers similar to the cancer cell line being tested, but without signs of cancer.
In vivo, compounds are evaluated in flank xenograft models established from sensitive SCLC (H889) and hematologic (RS4;11) cell lines, or using other tumor-forming cancer cell lines, according to what type of cancer is of particular interest to the user. When dosed orally or intravenously, compounds induce rapid and complete tumor responses (CR) that are durable for several weeks after the end of treatment in all animals bearing H889 (SCLC) or RS4;11 (ALL) tumors. Similar treatment of mice bearing H146 SCLC tumors can induce rapid regressions in the animals.
4
Ovarian Cancer Cell Line Cultivation and Characterization
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The human epithelial ovarian cancer cells A2780 and A2780CIS were purchased from the European Collection of Cell Cultures (ECACC). A2780CP20 cells were kindly gifted by Dr. Anil K. Sood (MD Anderson Cancer Center, Houston, TX) (16 (link)). High-grade serous ovarian cancer (HGSOC) cells OV-90 and OVCAR3 were purchased from ATCC (Chicago, IL). Cisplatin-resistant OV90CIS and OVCAR3CIS were generated by sequential addition of increasing concentrations of cisplatin to the parental cell lines (17 ). The chemosensitivity of the generated cell lines was assessed by dose-response experiments with cisplatin. The IC50 of this panel of cells has been reported (17 ). A2780, A2780CP20, and A2780CIS cells were maintained in RPMI-1640 (HyClone, Logan, UT), and OVCAR3 and OVCAR3CIS were maintained in RPMI-1640 supplemented with 0.01 mg/mL insulin (Sigma-Aldrich, St Louis, MO). OV-90 and OV-90CIS cells were cultivated on a 1:1 (v/v) ratio of M199 media and MCDB 105 media. All media was supplemented with 10% FBS and 1% antibiotics. For experiments, all cells were kept at 37°C and 5% CO2 atmosphere. Experiments were performed at 60–80% confluency.
5
Culturing Ovarian Cancer Cell Lines
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Ovarian cancer cell lines SKOV-3 and OVCAR-3 were purchased from American Type Culture Collection (ATCC) and cultured in RPMI culture medium containing 20% fetal bovine serum (FBS), 2 mM L-glutamine and 100 U/mL Penicillin-Streptomycin. OVCAR-3 cell culture cell culture medium was additionally supplemented with 0.01 mg/mL bovine insulin (cat# I6634, Sigma-Aldrich, St. Louis, MO, USA. Both cell lines were grown in the logarithmic phase at 37 °C in a 5%-CO2-humidified air.
6
Propagation and Tumorigenicity of OC Cell Lines
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Human OC cell lines (OVCAR-3 and A2780) and the mouse OC cell line (ID8) were purchased from the Committee on Type Culture Collection of the Chinese Academy of Sciences. OVCAR-3 cells were maintained in McCoy's 5A (Sigma), containing1% human insulin and 15% fetal calf serum (FCS). A2780 and ID8 cells were cultured with RPMI-1640 (Gibco) and DMEM (Gibco), respectively, supplemented with 10% FCS. Penicillin (100 IU mL−1) and gentamicin (40 IU mL−1) were added to the mediums for these three cells. All cells were cultured in a humidified incubator at 37 °C with 5% CO2.
Specific pathogen-free, 6–8-week-old female C57BL/6 littermate mice (20 ± 2 g) (Dossy Experimental Animals Co. LTD Chengdu, China) were used for tumorigenicity studies. Animal studies were carried out following protocols approved by the Ethics Committee of Sichuan Cancer Hospital (SCCHEC-04-2019-004).
Specific pathogen-free, 6–8-week-old female C57BL/6 littermate mice (20 ± 2 g) (Dossy Experimental Animals Co. LTD Chengdu, China) were used for tumorigenicity studies. Animal studies were carried out following protocols approved by the Ethics Committee of Sichuan Cancer Hospital (SCCHEC-04-2019-004).
7
Cell Line Maintenance Protocol
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OVCAR4, OVCAR3, 293T, and HeLa cells were purchased from the American Type Cell Culture (ATCC). A704 cells were obtained from Dr. Laura Banaszynski. OVCAR4 and OVCAR3 cells were maintained in RPMI (Sigma-Aldrich, R8758) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. HeLa, 293T and A704 cells were maintained in DMEM (Sigma-Aldrich, D5796) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. Fresh cell stocks were regularly replenished from the original ATCC-verified stocks and confirmed as mycoplasma-free every three months using a commercial testing kit.
8
Development of Cisplatin-Resistant Ovarian Cancer Cells
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Ovarian high-grade serous cancer cell lines, OC-3-VGH and OVCAR-3 cells, were obtained from Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan) and cell line authentication was performed by BCRC. OC-3-VGH cells were cultured in Dulbecco’s Modified Eagle Medium: Nutrient Mixture F-12 (DMEM/F12) with 1.5 g/L sodium bicarbonate and 10% fetal bovine serum, 100 IU/mL penicillin, 100 mg/mL streptomycin, and 0.4 mM L-glutamine (Sigma, St. Louis, MO, USA). OVCAR-3 cells were cultured in Roswell Park Memorial Institute (RPMI, Buffalo, NY, USA) 1640 with 1.5 g/L sodium bicarbonate, 2 mM L-glutamine, 2.5 g/L glucose, 10 mM HEPES, 1 mM sodium pyruvate, 10 ng/mL insulin and 20% fetal bovine serum, 100 IU/mL penicillin, 100 mg/mL streptomycin, and 0.4 mM L-glutamine (Sigma). Both cell lines were cultured in a humidified 95% atmosphere with 5% CO2 at 37 °C. We developed acquired cisplatin-resistant OV3-VGH cells using a stepwise increase in treatment concentrations (1 to 6 μM) with cisplatin (Sigma, St. Louis, MO, USA, Cat# 479306). This development period was carried out for about 2 months. Finally, the cisplatin-resistant cells (OC-3-VGH-resist) were kept in the DMEM/F12 medium with 6 μm cisplatin.
9
Culturing Ovarian Cancer Cell Lines
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Ovarian cancer cell lines, SKOV-3, HO9810, HO8910PM, and OVCAR3 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cell lines HO8910PM, HO9810, and OVCAR3 were cultured in RPMI-1640 culture medium, while SKOV-3 cells were cultured in McCoy’s 5A modified medium (Sigma-Aldrich, St. Louis MO, USA). All culture media were supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml of streptomycin.
10
Culturing Ovarian and Pancreatic Cancer Cells
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Human ovarian (OVCAR-3) and pancreatic (PANC-1) cancer cells were purchased from the American Type Culture Collection (ATCC, Rockville, MD) and maintained in exponential growth in a 5% CO2 humidified atmosphere at 37°C. OVCAR-3 cells were routinely grown in RPMI-1640 medium (Sigma, St. Louis, MO) supplemented with 20% FBS, L-glutamine (2 mM), antibiotics (100 IU penicillin/mL and 100 μg streptomycin/mL), insulin (50 ng/mL), and estradiol (1 nM). For OVCAR-3 cell proliferation assays, the medium used was identical, except for the omission of estradiol. PANC-1 cells were maintained in DMEM-high glucose (Invitrogen, Burlington, ON, Canada) containing L-glutamine (2 mM) and antibiotics (100 IU penicillin/mL and 100 μg streptomycin/mL), and supplemented with 10% FBS. For PANC-1 cell proliferation assays, the latter maintenance medium was replaced with DMEM/F12 containing the same supplements.
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