Kuramochi cell line

Manufactured by Japanese Collection of Research Bioresources Cell Bank

Sourced in Japan

The Kuramochi cell line is a well-characterized cell line derived from a human ovarian carcinoma. It is maintained and distributed by the Japanese Collection of Research Bioresources Cell Bank. The Kuramochi cell line is a valuable tool for research on ovarian cancer biology and the development of potential therapeutic interventions.

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KURAMOCHI (12 citations) KURAMOCHI cells (4 citations)

9 protocols using kuramochi cell line

Ovarian Cancer Cell Line Cultivation

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The NIH-OVCAR-3 cell line was obtained from ATCC (cod. HTB-161; American Type Culture Collection; Manassas, VA, USA). The OAW42 cell line was obtained from ECACC (cod. 85073102; European Collection of Authenticated Cell Cultures; Porton Down, Salisbury, UK). The KURAMOCHI cell line was obtained from JCRB Cell Bank (cod. JCRB0098; Japanese Collection of Research Bioresources Cell Bank; Japan). The OVCAR3 and KURAMOCHI human ovarian cancer cell lines were cultured in RPMI 1640 medium (cod. R8758; Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, cod. ECS0180L; Euroclone, Milan, Italy), 1% glutamine (cod. G7513; Sigma-Aldrich), and 1% penicillin/streptomycin (PES, cod. P0781; Sigma-Aldrich).
The OAW42 human ovarian cancer cell line was cultured in Minimum Essential Medium (MEM) (cod. M2279; Sigma-Aldrich) supplemented with 10% heat-inactivated FBS, 1% glutamine, 1% non-essential amino acids (cod. M7145; Sigma-Aldrich), and 1% PES.
The OVCAR3-GFP-LC3 cells were established in our laboratory [15 (link)] and maintained in culture conditions as the parental cell line.
All the cell lines were maintained under standard culture conditions (37 °C, 5% CO₂).

Esposito A., Ferraresi A., Salwa A., Vidoni C., Dhanasekaran D.N, & Isidoro C. (2022). Resveratrol Contrasts IL-6 Pro-Growth Effects and Promotes Autophagy-Mediated Cancer Cell Dormancy in 3D Ovarian Cancer: Role of miR-1305 and of Its Target ARH-I. Cancers, 14(9), 2142.

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Genetic Mouse Model Cell Lines for Ovarian Cancer Research

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The K-ras/PTEN cell line were established by us from ovarian tumors generated using a genetic mouse model.^{17 (link)} The SKOV3ip1 and HeyA8 cell lines were provided by Dr. Gordon Mills (MD Anderson Cancer Center, Houston, TX). The IOSE 397 cell line was kindly shared by Dr. Nelly Auersperg (University of British Columbia, Canada). The IGROV1 and Ovcar-5 cell lines were purchased from American Type Culture Collection (ATCC). The Kuramochi cell line was purchased from the Japanese Collection of Research Bioresources Cell Bank. Cell lines were validated by short tandem repeat (STR) DNA fingerprinting using the AMPF’STR Identifier kit (Applied Biosystems) and compared with ATCC and University of Texas MD Anderson Cancer Center fingerprints. Metformin obtained from Sigma-Aldrich (St Louis, MO). The Cdk4, cyclin D1, AMPK, EGFR, ErbB4, PDGFRα, FABP4 and FASN antibodies were from Cell Signaling Technologies (Beverly, MA) and the cyclin D1 antibody used for immunohistochemistry was from Novus Biologicals (Littleton, CO). The LKB1 and ACC antibodies were from Millipore (Billerica, MA), PARP 1/2 was from Santa Cruz Biotechnology (Santa Cruz, CA), FASN was from ATLAS (Stockholm, Sweden), phosphorylated RON was from R&D Systems (Minneapolis, MN), and RON was from Epitomics (Burlingame, CA).

LENGYEL E., LITCHFIELD L.M., MITRA A.K., NIEMAN K.M., MUKHERJEE A., ZHANG Y., JOHNSON A., BRADARIC M., LEE W, & ROMERO I.L. (2014). Metformin inhibits ovarian cancer growth and increases sensitivity to paclitaxel in mouse models. American journal of obstetrics and gynecology, 212(4), 479.e1-479.e10.

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Cell Line Authentication Protocols

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OVCAR-5, OVCAR-8, and CAOV-3 were purchased from American Type Culture Collection. SKOV3ip1 and HEYA8 were received from Dr. Gordon B. Mills (MD Anderson Cancer Center). The KURAMOCHI cell line was obtained from the Japanese Collection of Research Bioresources Cell Bank and the TYKNU cell line was obtained from Osaka University Graduate School of Medicine. The cell lines were validated by short tandem repeat DNA fingerprinting using the AmpFℓSTR Identifier kit (Applied Biosystems, Carlsbad, CA) and compared with known American Type Culture Collection fingerprints, the Cell Line Integrated Molecular Authentication database (CLIMA), and the University of Texas, MD Anderson Cancer Center fingerprint database.

Ladanyi A., Mukherjee A., Kenny H.A., Johnson A., Mitra A.K., Sundaresan S., Nieman K.M., Pascual G., Benitah S.A., Montag A., Yamada S.D., Abumrad N.A, & Lengyel E. (2018). Adipocyte-induced CD36 expression drives ovarian cancer progression and metastasis. Oncogene, 37(17), 2285-2301.

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Kuramochi Cell Line Wnt Pathway Modulation

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The Kuramochi cell line (RRID:CVCL_1345) was purchased from JCRB Cell Bank and cultured in RPMI 1640 medium (SH30027, HyClone) supplemented with 10% FBS, 50 units/ml penicillin, and 50 units/ml streptomycin (15140122, Gibco). Ascitic spheroids were cultured in MCDB/DMEM (1:1) medium (10372019 and 31966047, Gibco) supplemented with 10% FBS, 50 units/ml penicillin, and 50 units/ml streptomycin on ultra-low attachment plates (Corning). Cells and spheroids were stimulated with recombinant WNT proteins (645-WN-010, 5036-WN-CF; RnD Systems) at a concentration of 200 ng/ml or by WNT conditioned medium (CM) (control, WNT3A or WNT5A), produced by rat L-fibroblasts (CRL-2648, CRL-2647, CRL-2814; ATCC), at a ratio of 1:4 with complete culture medium. The Porcupine inhibitor LGK974 (974-02; Stem RD) was used at a 0.1 μM concentration while CK1 inhibitors PF670462 [sc-204180A; Santa Cruz Biotechnology (SCBT)] and D4476 (218696; Calbiochem) were used in 5 μM final concentration. If not stated otherwise, cells were pretreated with either LGK974 (+), or DMSO (-) for 24-48 h and treated with recombinant human (rh) WNTs/WNT CM overnight before being used for assays or WB.

Kotrbová A., Ovesná P., Gybel' T., Radaszkiewicz T., Bednaříková M., Hausnerová J., Jandáková E., Minář L., Crha I., Weinberger V., Záveský L., Bryja V, & Pospíchalová V. (2020). WNT signaling inducing activity in ascites predicts poor outcome in ovarian cancer. Theranostics, 10(2), 537-552.

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Comprehensive Ovarian Cancer Cell Lines

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We used 20 different human ovarian cancer cell lines, including Type I and Type II ovarian cancer cells (see Table S3 for details). The OVCAR8, PEO4, and A2780 cells were provided by Dr. S. Murphy (Duke University, Durham, NC, USA). The OVCAR3, OVCAR5, OVCAR7, OVCAR10, OVSAHO, and ES2 cell lines were provided by Dr. V. Shridhar (Mayo Clinic, Rochester, MN, USA). The Pt152 and Pt486 cells were provided by Dr. R. J. Buckanovich (University of Pittsburgh, Pittsburgh, PA, USA). The Kuramochi cell line was purchased from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). The CAOV3, SKOV3, NIH-OVCAR3, TOV112D (also known as TOV21D), HEY, and TOV21G cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The COV362 and PEO1 cells were purchased from the European Collection of Authenticated Cell Cultures (Millipore Sigma, Burlington, MA, USA).
All cell lines were cultivated in RPMI-1640 medium (Corning, Tewksbury, MA, USA) supplemented with 5% FBS (Thermo Fisher Scientific, Waltham, MA, USA) and 100 U/mL penicillin/100 μg/mL streptomycin (Corning, Tewksbury, MA, USA). The PEO1 cells were cultivated in medium with the addition of 1 mM sodium pyruvate (Corning, Tewksbury, MA, USA). The cells were tested for mycoplasma monthly.

Chesnokov M.S., Yadav A, & Chefetz I. (2022). Optimized Transcriptional Signature for Evaluation of MEK/ERK Pathway Baseline Activity and Long-Term Modulations in Ovarian Cancer. International Journal of Molecular Sciences, 23(21), 13365.

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Comprehensive Cell Line Protocol

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OVCAR3, A549, NIH-H23, HEK-293T, K562 and THP1 cells were purchased from ATCC; OVCAR4, OVCAR5 and OVACR8 cells were procured from National Cancer Institute; Kuramochi cell line was procured from JCRB Cell Bank. Human OSE cells were purchased from ScienCell Research Laboratories (no. 7310). We transduced OVCAR3 tumour cells with a luciferase-expressing vector (OVCAR3^luci). To make OVCAR3 cells as target of NY-ESO-1-specific T cells, OVCAR3^luci cells were transduced to express NY-ESO-1 (OVCAR3^luci-NY-ESO-1). Tumour-sorted CD45⁻EpCAM⁺ primary HGSOC cells were cultured continuously in R10 medium (RPMI-1640, 10% FBS, penicillin (100 IU ml⁻¹), streptomycin (100 μg ml⁻¹), l-glutamine (2 mM), sodium pyruvate (0.5 mM)) (ThermoScientific) until they adhere and grow similar to a cell line. All cell lines, except OSE, were routinely cultured in R10 medium. OSE cells were routinely cultured in recommended complete medium, purchased from ScienCell Research Laboratories. Cell lines routinely tested for negative mycoplasma contamination.

Biswas S., Mandal G., Payne K.K., Anadon C.M., Gatenbee C.D., Chaurio R.A., Costich T.L., Moran C., Harro C.M., Rigolizzo K.E., Mine J.A., Trillo-Tinoco J., Sasamoto N., Terry K.L., Marchion D., Buras A., Wenham R.M., Yu X., Townsend M.K., Tworoger S.S., Rodriguez P.C., Anderson A.R, & Conejo-Garcia J.R. (2021). IgA transcytosis and antigen recognition govern ovarian cancer immunity. Nature, 591(7850), 464-470.

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Cell Line Authentication and Maintenance

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OC cell lines OVCAR3 and A2780 were obtained from the American Type Culture Collection (ATCC, Manassas, VA); COV362 was obtained from Sigma. These cell lines were authenticated by short tandem repeat (STR) analysis in 2017 (IDEXX BioAnalytics, Columbia, MO). Kuramochi cell line was obtained from the Japanese Collection of Research Bioresources and authenticated by short tandem repeat (STR) analysis in 2018 (Clinical Molecular Oncology Laboratory, University of Kansas Cancer Center, KS). Cells were cultured as we have described previously (16 (link)). Cell lines were tested for mycoplasma contamination (Lonza) every 6 months. See Supplementary Material (SM) for more information.

Zong X., Wang W., Ozes A., Fang F., Sandusky G.E, & Nephew K.P. (2020). EZH2-mediated Downregulation of the Tumor Suppressor DAB2IP Maintains Ovarian Cancer Stem Cells. Cancer research, 80(20), 4371-4385.

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Cell Line Authentication Protocols

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Comprehensive Ovarian Cancer Cell Line Repository

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We used 20 different human ovarian cancer cell lines, including Type I and Type II ovarian cancer cells (see Table S1 for details). OVCAR8, PEO4, and A2780 cells were provided by Dr. S. Murphy (Duke University, Durham, NC, USA). OVCAR3, OVCAR5, OVCAR7, OVCAR10, OVSAHO and ES2 cell lines were provided by Dr. V. Shridhar (Mayo Clinic, Rochester, MN, USA). Pt152 and Pt486 cells were provided by Dr. R. J.
Buckanovich (University of Pittsburgh, Pittsburgh, PA, USA). The Kuramochi cell line was purchased from the Japanese Collection of Research Bioresources Cell Bank (Japan). CAOV3, SKOV3, NIH-OVCAR3, TOV112D (also known as TOV21D), HEY, and TOV21G cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). COV362 and PEO1 cells were purchased from European Collection of Authenticated Cell Cultures (Millipore Sigma, Burlington, MA, USA).
All cell lines were cultivated in RPMI-1640 medium (Corning, Tewksbury, MA, USA) supplemented with 5% FBS (Thermo Fisher Scientific, Waltham, MA, USA) and 100 U/mL penicillin/100 μg/mL streptomycin (Corning, Tewksbury, MA, USA). PEO1 cells were cultivated in medium with the addition of 1 mM sodium pyruvate (Corning, Tewksbury, MA, USA). Cells were tested for mycoplasma monthly.

Chesnokov M.S., Yadav A.K., & Chefetz I. (2022). Optimized transcriptional signature for evaluation of MEK/ERK pathway baseline activity and long-term modulations in ovarian cancer.

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