Ovcar3

Manufactured by Procell

Sourced in China

OVCAR3 is a well-established human ovarian carcinoma cell line derived from the ascites of a patient with papillary cystadenocarcinoma of the ovary. It serves as a valuable model for studying ovarian cancer biology and is widely used in cancer research.

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12 protocols using ovcar3

ACSM3 Regulation in Ovarian Cancer Cells

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OV-90, SK-OV-3, OVCAR-3, and A2780 cells were purchased from Procell Life Science&Technology Co., Ltd (Wuhan, China). OV-90 and OVCAR-3 cells were maintained in a specific medium (Procell). SK-OV-3 cells were cultured in McCoy’s 5A medium (Procell). A2780 cells were cultured in DEME (Gibco Life Technologies, NY, USA). All the medium was supplemented with 10% fetal bovine serum (FBS). And cells were incubated in 5% CO₂ at 37°C.
Lentiviral vectors expressing small hairpin RNA (shRNA) targeting ACSM3 were named Lv-shRNA1-ACSM3 or Lv-shRNA2-ACSM3. The complementary cDNAs of ACSM3 were synthesized and the lentiviral overexpressed vectors pcDNA3.1 (GenScript, Nanjing, China) of ACSM3 were constructed as Lv-ACSM3. The shRNA sequences were listed as follow. shRNA1-ACSM3: CGATGTTAAGATTGTAGATGT. shRNA2-ACSM3: GCTTGTACAGAATGATATAAC. The Lv-shRNA1-ACSM3 or the Lv-shRNA2-ACSM3 were infected into OV-90 cells. The Lv-ACSM3 was infected into A2780 or SK-OV-3 cells. The overexpressed ITG β1 (Integrin β1) vectors were transfected into A2780 cells by Lipofectamine 3000 (Invitrogen, Carlsbad, California, USA) according to the manufacturer’s instructions.

Yan L., He Z., Li W., Liu N, & Gao S. (2021). The Overexpression of Acyl-CoA Medium-Chain Synthetase-3 (ACSM3) Suppresses the Ovarian Cancer Progression via the Inhibition of Integrin β1/AKT Signaling Pathway. Frontiers in Oncology, 11, 644840.

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Inhibition of PHGDH, ROS, and PARP in Cancer Cells

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The PHGDH inhibitor CBR-5884, the ROS inhibitor N-acetylcysteine, and the PARP inhibitor olaparib were purchased from MedChemExpress. DMSO was used as control group in cell lines. Through literature reviewing, we used 5 mM NAC treated for 4 h to study its effect [36 (link), 37 (link)]. The cell lines (A2780, OVCAR3, and ES-2) were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China). A2780 and OVCAR3 were cultured with RPMI 1640 medium (Procell, Wuhan, China) containing 10% FBS (Procell, Wuhan, China), while ES-2 was cultured used McCoy's 5A (Procell, Wuhan, China) containing 10% FBS (Procell, Wuhan, China) in incubator. The incubator purchased from PUHE Biotechnology Co., Ltd. (Wuxi, China) was set at 37 °C, 5% CO2, and 1% O2 with enough humidity.

Zhang X., Sun M., Jiao Y., Lin B, & Yang Q. (2022). PHGDH Inhibitor CBR-5884 Inhibits Epithelial Ovarian Cancer Progression via ROS/Wnt/β-Catenin Pathway and Plays a Synergistic Role with PARP Inhibitor Olaparib. Oxidative Medicine and Cellular Longevity, 2022, 9029544.

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Ovarian Cancer Cell Culture Protocol

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Four OC cell lines (OVCAR3, SKOV3, A2780, and HO-8910) and human ovarian immortalized nontumorigenic ovarian surface epithelial cells (IOSE) were acquired from Procell Life Science Co., Ltd.,China and cultured in Dulbecco’s modified Eagle medium (DMEM) (TransGen Biotech Co., Ltd., Beijing, China) containing 10% fetal bovine serum (Gibco Company, NY, USA) and 1% penicillin/streptomycin (Sigma-Aldrich Chemical Company., MO, USA). All the cells were incubated in humidified chambers [21 (link)].

Wang Y., Li L., Zhang X, & Zhao X. (2022). Long non-coding RNA OIP5-AS1 suppresses microRNA-92a to augment proliferation and metastasis of ovarian cancer cells through upregulating ITGA6. Journal of Ovarian Research, 15, 25.

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Culturing OC Cell Lines SKOV3 and OVCAR-3

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SKOV3 and OVCAR-3 human OC cell lines were purchased from Procell Life Science & Technology Co., Ltd (cat. nos. CL-0215 and CL-0178). Short tandem repeat (STR) analysis was performed to confirm cell line authentication. SKOV3 or OVCAR-3 cells were cultured in McCoy's 5A or RPMI-1640 culture (Cytiva) supplemented with 10% fetal bovine serum (FBS; Invitrogen; Thermo Fisher Scientific, Inc.), streptomycin (100 mg/ml; Invitrogen; Thermo Fisher Scientific, Inc.) and penicillin (100 U/ml; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C in a humidified atmosphere, containing 5% CO₂.

Zhu X., Chen X., Qiu L., Zhu J, & Wang J. (2022). Norcantharidin induces ferroptosis via the suppression of NRF2/HO-1 signaling in ovarian cancer cells. Oncology Letters, 24(4), 359.

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Ovarian Cancer Cell Line SNHG25 Knockdown

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The normal ovarian surface epithelial cell line (IOSE80) and the ovarian cancer cell lines, SKOV3, A2780, HEY and OVCAR3, were purchased from Procell Life Science & Technology. Cells were cultured in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Gibco; Thermo Fisher Scientific, Inc.) at 37 °C in a humidified atmosphere and 5% CO₂.
For knockdown of SNHG25, A2780 and OVCAR3 cells were transfected with a lentiviral vector encoding shRNA targeting SNHG25 (5'-3': GGATGTCATCGTCCTTGCT) (LV-KD) or an empty vector as a negative control (LV-NC) (Genepharma, Shanghai, China) using polybrene (5.0 µg/mL). 24h after infection, cells were selected with 0.2 mg/mL puromycin. SNHG25 knockdown efficiency was confirmed with quantitative real-time polymerase chain reaction (qRT-PCR).

Liu Y., Xu B., Liu M., Qiao H., Zhang S., Qiu J, & Ying X. (2021). Long non-coding RNA SNHG25 promotes epithelial ovarian cancer progression by up-regulating COMP. Journal of Cancer, 12(6), 1660-1668.

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Ovarian Cell Lines Characterization

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The human ovarian cell lines A2780, SKOV3, OVCAR3, and OV90 and the normal human ovarian epithelial cell line IOSE80 were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China). All cell lines were cultured in DMEM medium (Servicebio, Wuhan, China) containing 10% FBS (Solarbio, Beijing, China) and stored in a humidified incubator at 37 °C with 5% CO₂. All the cell lines used in this study were re-authenticated by short tandem repeat (STR) profiling and tested for mycoplasma contamination in June 2021.

Lv W., Jia Y., Wang J., Duan Y., Wang X., Liu T., Hao S, & Liu L. (2022). Long non-coding RNA SNHG10 upregulates BIN1 to suppress the tumorigenesis and epithelial–mesenchymal transition of epithelial ovarian cancer via sponging miR-200a-3p. Cell Death Discovery, 8, 60.

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Overexpression and Knockdown of circ-ITCH in Ovarian Cancer Cells

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A2780 and OVCAR3 cell lines were purchased from Procell (Wuhan, China), and the ISOE80 cell line was obtained from the biotechnology company of Huzheng (Shanghai, China). All cells were cultured in Dulbecco’s modified eagle medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (FBS, Hyclone, South Logan, UT, USA) and 0.1% penicillin/streptomycin (Thermo Fisher Scientific) at 37 ℃ with 5% CO₂.
Overexpression vector of circ-ITCH was acquired by cloning the sequence of circ-ITCH into the pcDNA vector (RiboBio, Guangzhou, China). Small interfering RNAs against circ-ITCH (si-circ-ITCH) and CDH1 (si-CDH1) and their respective controls were synthesized by GenePharma (Shanghai, China). MiR-106a mimics, miR-control, miR-106a inhibitor, and anti-miR-control were synthesized by RiboBio. A2780 and OVCAR3 cells were transfected using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA).

Lin C., Xu X., Yang Q., Liang L, & Qiao S. (2020). Circular RNA ITCH suppresses proliferation, invasion, and glycolysis of ovarian cancer cells by up-regulating CDH1 via sponging miR-106a. Cancer Cell International, 20, 336.

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Ovarian Cancer Cell Culture Protocol

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The human OC cell lines OVCAR3 were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China). Cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco, NY, USA) containing 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin. The cells were maintained in an incubator at 37°C with 5% CO₂. Sodium oleate and sodium palmitate were obtained from Kunchuang Biotechnology (Xi’an, China).

Zhang L., Zhao X., Chu H., Zhao H., Lai X., Li J, & Lv T. (2022). Serum Free Fatty Acids and G-Coupled Protein Receptors Are Associated With the Prognosis of Epithelial Ovarian Cancer. Frontiers in Oncology, 12, 777367.

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Cell Culture of Human Ovarian Cell Lines

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The human EOC cell lines OVCAR-3, Caov-3, ES-2 and A2780 were purchased from the Chinese Academy of Sciences Cell Bank (Shanghai, China). The human normal ovarian epithelial cell line HOSEpiC were purchased from the American Type Culture Collection (ATCC, USA). OVCAR-3, Caov-3, A2780, and HOSEpiC were cultured in RPMI 1640 medium (Procell, Wuhan, China), supplemented with 10% fetal bovine serum (FBS; Procell). ES-2 was cultured in McCoy’s 5 A medium (Procell), also supplemented with 10% FBS. All cells were maintained under controlled conditions at a temperature of 37°C in an atmosphere containing 5% CO₂. Regular monitoring for mycoplasma contamination was performed using PCR.

Wang J., Zheng F., Wang D, & Yang Q. (2024). Regulation of ULK1 by WTAP/IGF2BP3 axis enhances mitophagy and progression in epithelial ovarian cancer. Cell Death & Disease, 15(1), 97.

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Ovarian Cell Lines Cultivation

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The normal human ovarian epithelial cell line (IOSE80) was purchased from Shanghai Fuheng Biotechnology (Shanghai, China). The human ovarian cancer cell lines (SKOV3, OVCAR3, ES2, A2780) were kindly provided by Procell Life Science & Technology Co., Ltd (Wuhan, China). Cells were cultured in the appropriate culture medium according to the instructions and were cultured at 37℃ in a humidified atmosphere with 5% CO₂. Human cells were short tandem repeat (STR)-profiled, used between passages 3 and 20, and regularly examined for mycoplasma.

Duan Y., Yu J., Chen M., Lu Q., Ning F., Gan X., Liu H., Ye Y., Lu S, & Lash G.E. (2023). Knockdown of heat shock protein family D member 1 (HSPD1) promotes proliferation and migration of ovarian cancer cells via disrupting the stability of mitochondrial 3-oxoacyl-ACP synthase (OXSM). Journal of Ovarian Research, 16, 81.

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