Cov362

Manufactured by Merck Group

Sourced in United States

COV362 is a laboratory equipment product manufactured by Merck Group. It is designed for analytical applications. The core function of COV362 is to provide accurate and reliable measurements for research and testing purposes.

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Lab products found in correlation

17 protocols using cov362

Ovarian Cancer Cell Culture Protocol

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We utilized the high grade serous and clear cell carcinoma ovarian cancer cell lines: SKOV3, ATCC (ATCC® #HTB-77™); COV362.4, Sigma Aldrich (Sigma #07071904) and OVCAR4, National Cancer Institute (RRID:CVCL_1627). Cells were maintained in: SKOV3 Dulbecco’s Modified Eagle Medium (DMEM)/Ham’s F-12 (DMEM/F12) (Thermo Scientific, #11965118); COV362.4 High glucose Dulbecco’s Modified Eagle Medium (DMEM-HG); and OVCAR4 RPMI-1640 (with sodium bicarbonate; Sigma-Aldrich, #R0883). Media was supplemented with 10% Fetal Calf Serum (FCS) (Thermo Fisher, #16000044) and 1% Penicillin-Streptomycin (Thermo Scientific, #15240062) with all lines maintained at 37 °C with 5% CO₂. Cell lines were verified and authenticated by the Australian Genome Research Facility Ltd. Cell counts were conducted prior to experimentation by Trypan Blue viability staining using the Countess® II viable cell count system, cells routinely tested negative for mycoplasma. 3D spheroids were formed by seeding cells into Corning® spheroid Ultra-low adhesion microplates (Sigma-Aldrich #CLS4520) with spheroid integrity, size and shape monitored by live cell microscopy to ensure uniformity across experiments. KRT14 overexpressing and knockout cell lines (LC^OE/ LC^KO) were generated according to the previously described methods [11 (link)].

Karimnia N., Wilson A.L., Green E., Matthews A., Jobling T.W., Plebanski M., Bilandzic M, & Stephens A.N. (2021). Chemoresistance is mediated by ovarian cancer leader cells in vitro. Journal of Experimental & Clinical Cancer Research : CR, 40, 276.

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Culturing Ovarian Cancer Cell Lines

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The epithelial ovarian cancer line OVCAR-3 (herein referred to as OVCAR3, ATCC HTB-161) was cultured in RPMI-1640 (Lonza) containing 20% fetal bovine serum (FBS, Hyclone) and 1X antibiotic-antimycotic (1X AA, Gibco) (complete RPMI). The epithelial ovarian cancer line derived from metastatic ascites OV-90 (herein referred to as OV90, CRL-11732) was cultured in a 1:1 mixture of MCDB 105 medium (Sigma) and Medium 199 (Thermo) adjusted to pH of 7.0 with sodium hydroxide (Sigma) and final 20% FBS and 1X AA. The epithelial-endometroid ovarian cancer line COV362.4 (Sigma) was cultured in Dulbecco's Modified Eagles Medium (DMEM, Life Technologies) containing 10% FBS, 1X AA, 25 mM HEPES (Irvine Scientific), and 2 mM L-Glutamine (Fisher Scientific) (complete DMEM). The epithelial ovarian cancer line OVCAR-8 (herein referred to as OVCAR8) was a generous gift from Dr. Carlotta Glackin at City of Hope and was cultured in complete RPMI-1640. The epithelial ovarian cancer line SK-OV-3 (herein referred to as SKOV3, ATCC HTB-77) and the colon epithelial cancer line LS 174T (herein referred to as LS174T, ATCC CL-188) were cultured in complete DMEM. DU145-PSCA cells were described previously (22 (link)). All cells were cultured at 37°C with 5% CO₂.

Murad J.P., Kozlowska A.K., Lee H.J., Ramamurthy M., Chang W.C., Yazaki P., Colcher D., Shively J., Cristea M., Forman S.J, & Priceman S.J. (2018). Effective Targeting of TAG72+ Peritoneal Ovarian Tumors via Regional Delivery of CAR-Engineered T Cells. Frontiers in Immunology, 9, 2268.

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Ovarian Cancer Cell Lines Characterization

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SUN119, ES-2, OV90, CaOV3, TOV21G, and the patient-derived cell line A39 were from Seoul National University, Seoul, Korea. Immortalized normal ovarian epithelial cells (OSE), fallopian-tube-derived epithelial cells (FTE188), and SKOV3-ip cell lines have been previously described and used [47 (link)–49 (link)]. OVCAR4 cell line was obtained from Dr. Thomas Hamilton (Fox Chase Cancer Center, PA). Kuramochi, TYKNU, and OVSAHO cell lines were from the JCRB Cell Bank, Tokyo, Japan while COV362, OAW28 and COV318 cells were purchased from Sigma-Aldrich (St. Louis, MO). Cell-culture conditions and the construction of bioluminescent JLP-silenced SKOV3-ip^Luc-shJLP and non-effective scrambled shRNA-expressing SKOV3-ip^LucshSC cell lines are detailed in Supplementary Material S1. Transient and stable transfections were carried out using a Nucleofector II from Lonza (Allendale, NJ) as previously described [50 (link), 51 (link)].

Ha J.H., Yan M., Gomathinayagam R., Jayaraman M., Husain S., Liu J., Mukherjee P., Reddy E.P., Song Y.S, & Dhanasekaran D.N. (2016). Aberrant expression of JNK-associated leucine-zipper protein, JLP, promotes accelerated growth of ovarian cancer. Oncotarget, 7(45), 72845-72859.

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Cell Culture of Human Tumor Lines

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Human tumor cell lines (PSN1, HCC1806, PANC1, SW1990, HS766T, PANC0504, NCIH1792, NCIH23, NCIH1651, MDAMB436) were obtained from American Type Culture Collection (ATCC) and were cultured under ATCC recommended media and condition. COV362 was obtained from Sigma-Aldrich and was maintained in DMEM media with 10% FBS. All cells were incubated at 37 °C in a humidified atmosphere with 5% CO₂.

Ding X., Zhu Z., Lapek J., McMillan E.A., Zhang A., Chung C.Y., Dubbury S., Lapira J., Firdaus S., Kang X., Gao J., Oyer J., Chionis J., Rollins R.A., Li L., Niessen S., Bagrodia S., Zhang L, & VanArsdale T. (2022). PARP1-SNAI2 transcription axis drives resistance to PARP inhibitor, Talazoparib. Scientific Reports, 12, 12501.

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Cell Culture Conditions for Cancer Research

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RPE1-hTERT p53^−/− Cas9
(Zimmermann et al., 2018 (link)) and
293T cells were grown in DMEM supplemented with 10% FBS (Wisent
#080150), GlutaMAX (life Technologies #35050–061) and 1%
Pen/Strep (Wisent #450–201-EL). UWB1.28, SKOV3 and OV90 cells
were purchased from ATCC. UWB1.28 cells were grown in RPMI: MEGM (1:1)
supplemented with 3% FBS and 1% Pen/Strep. SKOV3 cells were grown in
McCoys 5a supplemented with 10% FBS and 1% Pen/Strep. OV90 cells were
grown in MCDB 105: Medium 99 (1:1) supplemented with 1% sodium
bicarbonate (0.75 g/L), 15% FBS and 1% Pen/Step. JHOS2 were purchased
from Riken BRC and grown in DMEM: F-12(1:1) supplemented with 10%FBS,
0.1mM NEAA. SUM149PT cells were purchased from Asterand BioScience and
grown in a DMEM/F12 medium mixture supplemented with 5% FBS, 1%
Pen/Strep, 1 μg/mL hydrocortisone and 5 μg/mL insulin.
COV362 and COV644 were purchased from Sigma and grown in DMEM
supplemented with 10% FBS and 1% Pen/Strep. RMUGS cells were purchased
from JCRC cell bank and grown in Ham’s 12 supplemented with 10%
FBS and 1% Pen/Strep. Wild-type and
BRCA2^−/−DLD-1 cells were purchased from Horizon and maintained in RPMI medium
supplemented with 10% FBS, 1% Pen/Strep and 2 mM sodium pyruvate. All
cell lines were grown at 37 °C and 5% CO2.

Álvarez-Quilón A., Wojtaszek J.L., Mathieu M.C., Patel T., Appel C.D., Hustedt N., Rossi S.E., Wallace B.D., Setiaputra D., Adam S., Ohashi Y., Melo H., Cho T., Gervais C., Muñoz I.M., Grazzini E., Young J.T., Rouse J., Zinda M., Williams R.S, & Durocher D. (2020). Endogenous DNA 3′ blocks are vulnerabilities for BRCA1 and BRCA2 deficiency and are reversed by the APE2 nuclease. Molecular cell, 78(6), 1152-1165.e8.

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Culturing Ovarian Cancer Cell Lines

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The human ovarian cancer cells COV318 and COV362 (Sigma, MO, USA) were grown at 37°C in Dulbecco's Modified Eagle Medium supplemented with 10% fetal bovine serum (FBS), L-glutamine, and pencillin/streptomycin in a humidified atmosphere containing 5% CO₂. Everolimus (RAD001) and NVP-BEZ235 were obtained from Selleckchem (Provided by Sapphire Biosciences, NSW, Australia). Short tandem repeat profiling and mycoplasma testing (MycoAlert^TM Plus Mycoplasma detection kit, Lonza, MD, USA) were conducted at regular intervals for the quality control of cell culture conditions and validation of these cell lines.

Bajwa P., Nagendra P.B., Nielsen S., Sahoo S.S., Bielanowicz A., Lombard J.M., Wilkinson E.J., Miller R.A, & Tanwar P.S. (2016). Age related increase in mTOR activity contributes to the pathological changes in ovarian surface epithelium. Oncotarget, 7(15), 19214-19227.

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Ovarian Cancer Cell Lines Authentication

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HeyA8, HeyA8-MDR, A2870IP2 (referred to as A2870 throughout), A2780CP20, COV362, SKOV3, SKOV3IP1, SKOV3TRIP2, and HI0-180 cells were obtained from Dr. Anil Sood (MD Anderson Cancer Center, Houston, TX) and have been described previously [46 (link)]. COV362 were obtained from Sigma-Aldrich (St. Louis, MO). All cell lines were authenticated by short tandem repeat DNA profiling prior to experimental studies. OVCAR4 cells were obtained from Charles River Laboratories (New York, NY). All cells were maintained in RPMI (Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS), with no antibiotics, and were passaged <20 times.

McQueeney K.E., Salamoun J.M., Burnett J.C., Barabutis N., Pekic P., Lewandowski S.L., Llaneza D.C., Cornelison R., Bai Y., Zhang Z.Y., Catravas J.D., Landen C.N., Wipf P., Lazo J.S, & Sharlow E.R. (2017). Targeting ovarian cancer and endothelium with an allosteric PTP4A3 phosphatase inhibitor. Oncotarget, 9(9), 8223-8240.

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Isolation and Characterization of Human ECFCs

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Human ECFCs were isolated from neonatal cord and peripheral blood and their distinct endothelial phenotypes were verified by flow cytometry as previously described (see Supporting Information Fig. S1) (Hofmann et al., 2009; 2012,; Reinisch and Strunk, 2009 (link, link, link); Reinisch et al., 2009 (link)). HUVECs were obtained from Lonza (Basel, Switzerland). ECFC and HUVECs were grown in endothelial growth medium-2 (EGM-2) (Lonza) containing 2% FBS and 1% penicillin/streptomycin/L-glutamine/heparin (Life Technologies, Carlsbad, CA, USA) and EGM-2 growth factor supplements (composed of bFGF, IGF-2, EGF, VEGF, ascorbic acid, hydrocortisone). Ovarian carcinoma cell lines OVCAR-3 (American Type Culture Collection, Manassas, VA, USA), OVCAR-5 (kindly provided by the Cell Culture Core, Vascular Biology Program, Boston Children's Hospital, Boston, MA, USA) and COV-362 (Sigma Aldrich, St. Louis, MO, USA) were grown in DMEM containing 10% FBS.

Hofmann N.A., Yang J., Trauger S.A., Nakayama H., Huang L., Strunk D., Moses M.A., Klagsbrun M., Bischoff J, & Graier W.F. (2015). The GPR 55 agonist, L-α-lysophosphatidylinositol, mediates ovarian carcinoma cell-induced angiogenesis. British Journal of Pharmacology, 172(16), 4107-4118.

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Culturing Human Ovarian Cancer Cell Lines

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The human, female ovarian carcinoma cell lines COV318, COV362 (Sigma), CAOV3 (ATCC) were cultured in DMEM, while OVCAR3 (ATCC), Kuramochi, OVSAHO, OVMANA and OVISE (JCRB Cell Bank) were cultured in RPMI. RMG1 (JCRB Cell Bank) were cultured in Hams-F12 media. All cell lines were grown with 10% fetal bovine serum, 100 U/ml penicillin, 100 U/ml streptomycin and 2 mM glutamine and were maintained at 37°C in a humidified 5% CO₂ atmosphere. OV56 (Sigma) were cultured in DMEM/F12 as above but supplemented with 10 μg/ml insulin, 0.5 μg/ml hydrocortisone and 5% fetal bovine serum. All lines were authenticated by the Molecular Biology Core Facility at the CRUK Manchester Institute using Promega Powerplex 21 System and periodically tested for mycoplasma.

Pillay N., Tighe A., Nelson L., Littler S., Coulson-Gilmer C., Bah N., Golder A., Bakker B., Spierings D.C., James D.I., Smith K.M., Jordan A.M., Morgan R.D., Ogilvie D.J., Foijer F., Jackson D.A, & Taylor S.S. (2019). DNA Replication Vulnerabilities Render Ovarian Cancer Cells Sensitive to Poly(ADP-Ribose) Glycohydrolase Inhibitors. Cancer Cell, 35(3), 519-533.e8.

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Cell Line Authentication and Validation

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All cell lines were grown in DMEM with 10% FBS along with 1% MEM vitamins, MEM nonessential amino acids and Penicillin-Streptomycin. CAOV3, HeyA8, OVCAR5 and OVCAR8 were obtained from Ernst Lengyel, University of Chicago and OVCAR4 from Joanna Burdette, University of Illinois at Chicago. OVSAHO was obtained from the Japanese Collection of Research Bioresources and SNU119 was from the Korean Cell Line Bank. COV362 (Sigma), and Kuramochi (Japanese Collection of Research Bioresources). All cell lines used were genetically validated and also confirmed to be mycoplasma free using respective services from Idexx BioResearch (Columbia, MO). The genetic validation was done using the CellCheck 16 (16 Marker STR Profile and Inter-species Contamination Test) and mycoplasma testing was done using Stat-Myco.

Haley J., Tomar S., Pulliam N., Xiong S., Perkins S.M., Karpf A.R., Mitra S., Nephew K.P, & Mitra A.K. (2016). Functional characterization of a panel of high-grade serous ovarian cancer cell lines as representative experimental models of the disease. Oncotarget, 7(22), 32810-32820.

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