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Ovcar 3 cells

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The OVCAR-3 cells are a well-characterized human ovarian adenocarcinoma cell line derived from the malignant ascites of a patient with progressive ovarian cancer. These cells are widely used in cancer research to study the biology and treatment of ovarian cancer.

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Lab products found in correlation

Rpmi 1640 medium, by Thermo Fisher Scientific (5 mentions) Rpmi 1640 medium, by American Type Culture Collection (3 mentions) Human synoviocyte growth medium, by Cell Applications (3 mentions) Hfls ra, by Cell Applications (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by American Type Culture Collection (2 mentions) Caov3, by American Type Culture Collection (2 mentions) Es 2 cells, by American Type Culture Collection (2 mentions) Cov362, by Merck Group (2 mentions) Cov318, by Merck Group (2 mentions) Rpmi medium, by Thermo Fisher Scientific (2 mentions) Ripa buffer, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Merck Group (2 mentions) Rpmi 1640, by Merck Group (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Lipofectamine, by Thermo Fisher Scientific (1 mentions) Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions) Hs27 cells, by American Type Culture Collection (1 mentions)

27 protocols using ovcar 3 cells

1

Ovarian Cancer Cell Culturing and Co-Culture

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The majority of the experiments described here used OVCAR-3 cells (ATCC, Manassas, VA), a human epithelial ovarian cancer cell line. OVCAR-3 cells were subcultured under standard cell culture conditions in RPMI medium supplemented with 20% fetal bovine serum, 0.01 mg/mL bovine insulin, and 1% penicillin–streptomycin. For experimentation, near confluent cells from stock T-75 flasks were detached with 0.25% trypsin–EDTA and subcultured onto 18 mm round glass coverslips at a 1:5 split ratio and grown for 5 days in standard culture conditions prior to experimentation. OVCAR-3 cells used here were passage number 35–50.
For co-culture experiments, Hs-27 cells (ATCC), a human fibroblast cell line, were seeded at equal densities and grown with OVCAR-3 cells on glass coverslips under similar conditions. Dulbecco’s Modified Eagle’s Medium with 10% fetal bovine serum was mixed at a 1:1 ratio with the RPMI medium described above for co-culturing Hs-27 and OVCAR-3 cells.
Orsinger G.V., Williams J.D, & Romanowski M. (2014). Focal Activation of Cells by Plasmon Resonance Assisted Optical Injection of Signaling Molecules. ACS Nano, 8(6), 6151-6162.
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2

Culturing Human Cell Lines

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Human epithelial ovary adenocarcinoma (OVCAR-3) human T-lymphocyte cells (Jurkat, E6–1) were purchased from American Type Culture Collection (ATCC). Jurkat cells were cultured in RPMI-1640 medium (ATCC) with 10% FBS, 0.01 mg/mL bovine insulin and 0.1 mg/mL penicillin/streptomycin. OVCAR-3 cells were cultured in RPMI-1640 medium from ATCC, with 20% FBS, 0.01 mg/mL bovine insulin, and 0.1 mg/mL penicillin/streptomycin. Human fibroblast-like synoviocytes rheumatoid arthritis (HFLS-RA) cells were purchased from Cell Applications, Inc., San Diego, CA, USA, and maintained in a human synoviocyte growth medium purchased from Cell Applications, Inc. All cell lines were maintained at 37 °C and 5% CO2.
Dahal A., Parajuli P., Singh S.S., Shrestha L., Sonju J.J., Shrestha P., Chatzistamou I, & Jois S. (2022). Targeting protein–protein interaction for immunomodulation: A sunflower trypsin inhibitor analog peptidomimetic suppresses RA progression in CIA model. Journal of pharmacological sciences, 149(3), 124-138.
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3

Generating CD20-positive Ovarian Cancer Cells

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Daudi and Ramos (used as target cells), and Jurkat cell lines were purchased from the American Type Culture Collection (ATCC). Cells were cultured in complete RPMI (cRPMI) medium consisting of RPMI (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), placed at 37°C in a humidified incubator containing 5% CO2 and passaged twice weekly. Jurkat-Luc effector cells were purchased from Promega Corporation, cultured in cRPMI and placed at 37°C in a humidified incubator containing 5% CO2. CA125-positive OVCAR3 cells (13 (link)) (Fig. 3E) were purchased from ATCC and grown in cRPMI. To generate CA125- and CD20-positive target cells, OVCAR3 cells were grown in cRPMI and transfected using Lipofectamine (Thermo Fisher Scientific, Inc.) with 10 µg CD20 cDNA expression plasmid (pUNO1-hCD20a; InvivoGen). Transfected cells were selected using 30 µg/ml blasticidin.
Grasso L., Kline J.B, & Nicolaides N.C. (2021). Block-Removed Immunoglobulin Technology to enhance rituximab effector function by counteracting CA125-mediated immunosuppression. Oncology Letters, 23(1), 2.
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4

Culturing Human Cell Lines

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Human epithelial ovary adenocarcinoma (OVCAR-3) human T-lymphocyte cells (Jurkat, E6–1) were purchased from American Type Culture Collection (ATCC). Jurkat cells were cultured in RPMI-1640 medium (ATCC) with 10% FBS, 0.01 mg/mL bovine insulin and 0.1 mg/mL penicillin/streptomycin. OVCAR-3 cells were cultured in RPMI-1640 medium from ATCC, with 20% FBS, 0.01 mg/mL bovine insulin, and 0.1 mg/mL penicillin/streptomycin. Human fibroblast-like synoviocytes rheumatoid arthritis (HFLS-RA) cells were purchased from Cell Applications, Inc., San Diego, CA, USA, and maintained in a human synoviocyte growth medium purchased from Cell Applications, Inc. All cell lines were maintained at 37 °C and 5% CO2.
Dahal A., Parajuli P., Singh S.S., Shrestha L., Sonju J.J., Shrestha P., Chatzistamou I, & Jois S. (2022). Targeting protein–protein interaction for immunomodulation: A sunflower trypsin inhibitor analog peptidomimetic suppresses RA progression in CIA model. Journal of pharmacological sciences, 149(3), 124-138.
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5

Cultivation of SKOV-3 and OVCAR-3 Cell Lines

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SKOV-3 cells were cultivated in McCoys medium (Biochrom AG, Berlin, Germany) containing 10% fetal bovine serum and 1% penicillin-streptomycin (Invitrogen, Karlsruhe, Germany) at 37°C in a 5% CO2 humidified atmosphere. OVCAR-3 cells purchased from ATCC (Manassas, Virginia, USA) were cultivated in RPMI medium (Invitrogen) containing 10% fetal bovine serum, 1% penicillin-streptomycin and 0.01 mg/ml insulin under conditions described above.
Mayer C., Darb-Esfahani S., Meyer A.S., Hübner K., Rom J., Sohn C., Braicu I., Sehouli J., Hänsch G.M, & Gaida M.M. (2016). Neutrophil Granulocytes in Ovarian Cancer - Induction of Epithelial-To-Mesenchymal-Transition and Tumor Cell Migration. Journal of Cancer, 7(5), 546-554.
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6

Ovarian Cancer Cell Culture Protocol

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Cells were cultured in 10 cm plates in a humidified atmosphere (5% CO2) at 37 °C. At 70–90% confluence, trypsin (0.25%)/EDTA solution (Corning) was used to detach the cells from the culture plate for passaging and used for further experiments until passage 20. OV81.2 and OV81.2-CP10 primary cell line models were generated as previously described [10 (link)]. A2780 and CP70 cells were obtained from Dr. Paul Modrich (Duke University). OVCAR3 cells were purchased from ATCC. OV81.2, OV81.2-CP10, A2780, CP70, ALDHhigh CP70, ALDHlow CP70, OVCAR8 and 293 T cells were cultured in DMEM medium supplemented with 10% FBS (Gibco) and 1% penicillin-streptomycin (PS) (Gibco). OVCAR3 and OVCAR3-CP38 cells were cultured in RPMI medium (Gibco) with 10% FBS and 1%PS. Phosphate-buffered saline (PBS) was purchased from Corning. Amiodarone, dronedarone, ibutilide and sotalol were purchased from SelleckChem. Dimethyl sulfoxide (DMSO) was purchased from Fisher Chemical.
Nagaraj A.B., Joseph P., Kovalenko O., Wang Q., Xu R, & DiFeo A. (2018). Evaluating class III antiarrhythmic agents as novel MYC targeting drugs in ovarian cancer. Gynecologic oncology, 151(3), 525-532.
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7

Ovarian Cancer Cell Line Characterization

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Human ovarian cancer cell line 200845 (link),46 (link) was provided by Dr. Francois X. Claret (MD Anderson Cancer Center). SKOV3 and PEO4 cell lines were provided by Dr. Thomas C. Hamilton (Fox Chase Cancer Center). A2780/CP70 cell line was provided by Dr. Paul Modrich (Duke University). OVCAR3 cells were purchased from ATCC (Manassas, VA). CP70 cells stably transfected with pcDNA3.1-His-DDB2 (CP70-DDB2-1B) were established as described previously12 . All cell lines were authenticated by STR profiling and tested for mycoplasma contamination. These cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 µg/mL streptomycin, and 100 units per mL penicillin. Doxycycline (Dox) was purchased from Sigma-Aldrich. Isopropyl β-d-1-thiogalactopyranoside (IPTG) was purchased from ThermoFisher. The ALDH1A1 selective inhibitor NCT-501 was provided by Dr. David Maloney. For treatment of in vitro-cultured cells, NCT-501 was dissolved in DMSO. For treatment of mice by intraperitoneal injection, NCT-501 was dissolved in 20% 2-hydroxypropryl-β-cyclodetrin (HPβCD) in saline.
Cui T., Srivastava A.K., Han C., Wu D., Wani N., Liu L., Gao Z., Qu M., Zou N., Zhang X., Yi P., Yu J., Bell E.H., Yang S.M., Maloney D.J., Zheng Y., Wani A.A, & Wang Q.E. (2018). DDB2 represses ovarian cancer cell dedifferentiation by suppressing ALDH1A1. Cell Death & Disease, 9(5), 561.
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8

Cell Culture Conditions for Cancer and Immune Research

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Human ovarian carcinoma cells (OVCAR-3) and human T-lymphocyte cells (Jurkat, E6-1) were purchased from the American Type Culture Collection (ATCC). OVCAR-3 cells were cultured in RPMI-1640 medium from ATCC, with 20% FBS, 0.01 mg/mL bovine insulin, and 0.1 mg/mL penicillin/streptomycin. Jurkat cells were cultured in RPMI-1640 medium (ATCC) with 10% FBS, 0.01 mg/mL bovine insulin and 0.1 mg/mL penicillin/streptomycin. Human fibroblast-like synoviocytes rheumatoid arthritis (HFLS-RA) cells were purchased from Cell Applications, Inc., San Diego, CA, USA, and maintained in human synoviocyte growth medium purchased from Cell Applications, Inc. OVCAR-3 cells and HFLS-RA cells were used as a model for cells expressing CD58 protein, and Jurkat cells were used as a model for human T cells.
Parajuli P., Sable R., Shrestha L., Dahal A., Gauthier T., Taneja V, & Jois S. (2020). Modulation of costimulatory signal from CD2-CD58 proteins by a grafted peptide. Chemical biology & drug design, 97(3), 607-627.
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9

Cell Culture and Lysis for Pulldown Assays

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Cells were maintained at 37 °C and 5% CO2. HeLa cells (ATCC CCL-2) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S). Capan-2 cells (ATCC HTB-80) were cultured in McCoy’s 5a supplemented with 10% FBS and 1% P/S. K562 and SKBR3 cells (ATCC CRL-3344 and HTB-30, respectively) were cultured in RPMI supplemented with 10% FBS and 1% P/S. OVCAR-3 cells (ATCC HTB-161) were cultured in RPMI supplemented with 20% FBS, 0.01 mg/mL bovine insulin, and 1% P/S. To prepare lysate for pulldowns, cells plated in T75 flasks (Thermo Fisher Scientific) were grown until ~70% confluency, washed three times with DPBS, then lysed in 500 µL of RIPA buffer (Thermo Fisher Scientific) supplemented with EDTA-free protease inhibitor cocktail (Roche) and 0.1% benzonase (Millipore Sigma). Lysates were stored at −80 °C prior to pulldown.
Malaker S.A., Riley N.M., Shon D.J., Pedram K., Krishnan V., Dorigo O, & Bertozzi C.R. (2022). Revealing the human mucinome. Nature Communications, 13, 3542.
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10

Feeder-independent Mouse Embryonic Stem Cells Culture

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Feeder-independent mouse ESCs59 (link) were cultured on gelatin (0.2%) coated plates in mESC self-renewal medium (DMEM with 15% knockout serum replacement (Gibco), 1% nonessential amino acids (Gibco), 1 mM sodium pyruvate (Invitrogen), 1% L-glutamine (Invitrogen), 0.1 mM ß-mercaptoethanol, 1% penicillin/streptomycin (Invitrogen) and 1,000 U ml−1 mouse leukaemia inhibitory factor. pMEFs were generated from 13.5 d.p.c C57BL/6 mouse embryos. NIH-3T3 cells were obtained from ATCC. pMEFs and NIH-3T3 cells were maintained in culture in DMEM supplemented with 10% foetal bovine serum (Lonza) and 1% penicillin/streptomycin. iPS were generated and cultivated according to standard conditions60 (link). OVCAR-3 cells (ATCC) were cultured in RPMI-1640 medium supplemented with 20% foetal bovine serum (Lonza), L-glutamine (2 mM), insulin (10 μg ml−1; I9278, Sigma) and 1% penicillin/streptomycin. SKOV-3 cells (ATCC) and U2OS cells (ATCC) were maintained in DMEM supplemented with 10% foetal bovine serum and 1% penicillin/streptomycin. To generate stable mESC lines, cells were transfected (TransIT-LT1; Mirus Bio LCC) with respective vectors and subjected to selection using G418 (Sigma, 300 μg μl−1) or puromycin (Sigma, 3 μg ml−1).
Scarola M., Comisso E., Pascolo R., Chiaradia R., Maria Marion R., Schneider C., Blasco M.A., Schoeftner S, & Benetti R. (2015). Epigenetic silencing of Oct4 by a complex containing SUV39H1 and Oct4 pseudogene lncRNA. Nature Communications, 6, 7631.
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