Hela cells were purchased from the ATCC (Manassas, VA, USA) and cultured in DMEM with 10% FBS and 1% penicillin/streptomycin. Hela-CD19 cells with stable expression of CD19 were maintained in DMEM with 10% FBS, puromycin and penicillin/streptomycin.as described in [33 (link)]. Human peripheral blood mononuclear cells (PBMC) were isolated from whole blood obtained in the Stanford Hospital Blood Center (Stanford, CA, USA) according to IRB-approved protocol using Ficoll-Paque solution (GE Healthcare, Chicago, IL, USA). HEK293FT cells from AlStem (Richmond, CA, USA) were cultured in DMEM containing 10% FBS and penicillin/streptomycin. SKOV-3 cell line was obtained from ATCC and cultured in RPMI plus 10% FBS and penicillin/streptomycin. Normal human keratinocytes were obtained from the Lonza company and cultured in keratinocyte medium (Lonza, Anaheim, CA, USA) according to the manufacturer’s protocol. BxPC3, PANC-1, A1847, A375, A549 and Hep-3 B were obtained from ATCC and cultured in DMEM with 10% FBS and penicillin/streptomycin. The cell lines were authenticated by flow cytometry in our laboratory, using cell-specific surface markers or by ATCC.
Skov3 cell line
Manufactured by American Type Culture Collection
Sourced in United States
The SKOV3 cell line is a human ovarian adenocarcinoma cell line derived from the ascites of a patient with a poorly differentiated papillary cystadenocarcinoma. It is a widely used in vitro model for ovarian cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Merck Group (4 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Streptomycin, by Thermo Fisher Scientific (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Merck Group (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions)
Gfr matrigel, by BD (2 mentions)
Ovtoko, by Japanese Collection of Research Bioresources Cell Bank (2 mentions)
Ovise, by Japanese Collection of Research Bioresources Cell Bank (2 mentions)
Tov21g, by Japanese Collection of Research Bioresources Cell Bank (2 mentions)
Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (2 mentions)
Ficoll paque solution, by GE Healthcare (1 mentions)
A1847, by American Type Culture Collection (1 mentions)
Hep3b, by American Type Culture Collection (1 mentions)
Hela cell, by American Type Culture Collection (1 mentions)
35 protocols using skov3 cell line
1
Cell Culture Protocol for Multiple Cell Lines
2
Ovarian Carcinoma Cell Line Cultivation
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The human ovarian carcinoma cell lines A2780 and SKOV‐3 were used in this study. A2780 cell line was procured from Sigma‐Aldrich, St Louis, MO, USA, and cultured in RPMI 1640 medium (Invitrogen, Carlsbad, CA, USA). SKOV‐3 cell line was procured from ATCC, Manassas, VA, USA, and cultured in McCoy's 5a medium (ATCC). All media were supplemented with 10% (v/v) heat‐inactivated fetal bovine serum (Biowest LLC, Riverside, MO, USA), 100 U·mL−1 penicillin, and 100 U·mL−1 streptomycin (Sigma‐Aldrich). For routine culture, cells were grown until reaching approximately 80% confluency and then plated for experiments.
3
Cytotoxicity Assays for Cancer Cell Lines
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A549 cell line (human lung cancer cell line), Hela cell line (human cervical carcinoma cell line), SK-OV-3 cell line (human ovarian cancer), NIC-H1975 cell (human lung cancer cell line), K562 cell line (human leukemic cell line), S180 cell line (mouse sarcoma cell line), and H22 (mouse hepatoma cell line) were obtained from the ATCC (Manassas, VA, USA). SB203580 or SP600125 was purchased from Dalian Meilun Biology Co., Ltd. (Dalian, China). RPMI1640 medium was purchased from Gibco BRL, Life Technologies (USA). Fetal bovine serum (FBS) was purchased from Hyclone Laboratories (Logan, Utah, USA). The antibodies were purchased from Cell Signaling Technology. FITC-conjugated, Annexin V, and PI were purchased from BD Bioscience.
4
Nanoparticle Delivery of Doxorubicin Derivative
5
Establishment and Characterization of SKOV-3 Cell Line
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The SKOV-3 cell line (ATCC, Manassas VA, USA; Cat. No. HTB-77) was originally established from the malignant ascitic fluid of a patient with progressive ovarian carcinoma at Memorial Sloan-Kettering Cancer Center. SKOV-3 cells are capable of generating tumors in the peritoneal cavity of nude mice but are unable to form cell aggregates. Primary cell cultures (INCan017 and INCan019) were obtained from the ascitic fluid from the 2 abovementioned patients (see below).
6
Silencing RAD51AP1 in SKOV3 Ovarian Cancer Cells
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The SKOV3 cell line (ATCC, USA) was used as an in vitro model for OvCa. Cells were grown in Roswell Park Memorial Institute (RPMI) media supplemented with 10% FBS (Gibco) and 1% penicillin/streptomycin (Gibco). Cell lines were cultured at 37 °C, in 5% CO2 environment. siRNA targeted to RAD51AP1 (SMARTpool: ON-TARGETplus, Dharmacon, CO, USA) was used to suppress RAD51AP1 gene/protein expression in SKOV3 cells, as previously described [3 (link)].
7
Knockdown of WNK1, MEKK2, MEK5, and ERK5 in Ovarian Cancer Cell Lines
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SKOV3 cell line was obtained from the ATCC. OVCAR8, A2780 and IGROV1 ovarian cancer cell lines were from Pharmamar (León, Spain). Their authenticity was checked by STR profiling at the University Hospital of Salamanca. All ovarian cancer cell lines were cultured at 37°C in a humidified atmosphere in the presence of 5% CO2 and 95% air. Cells were grown in DMEM medium containing high glucose (4.5 g/L), l ‐glutamine 4 mM and l ‐pyruvate 5 mM (SKOV3 and OVCAR8) or RPMI medium containing l ‐glutamine 4 mM (A2780 and IGROV1). The medium was supplemented with 10% FBS and antibiotics (penicillin 100 U/mL and streptomycin 100 μg/mL). Cell culture media, FBS and penicillin–streptomycin were purchased from GIBCO BRL (Gaithersburg, MD, USA). Cells were maintained in culture for up to 4 months and mycoplasma testing was periodically performed.
Knockdown of WNK1, MEKK2, MEK5 and ERK5 were performed as described27 by infection of cell lines with TRC lentiviral pLKO vectors containing human WNK1 (gene set RHS4533‐EG65125), MAP3K2 (gene set RHS4533‐EG10746), MAP2K5 (gene set RHS4533‐EG5607) or MAPK7 (gene set RHS4533‐EG5598) sequences purchased from Thermo Fisher Scientific (Waltham, MA, USA).
Knockdown of WNK1, MEKK2, MEK5 and ERK5 were performed as described
8
Evaluating CC-I Cytotoxicity on Ovarian Cancer
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Example 9
Human ovarian adenocarcinoma SKOV-3 cell lines were ordered from American Type Culture Collection (ATCC, Manassas, Va.), and maintained in McCoy's 5A media with 100 U/ml penicillin, 100 μg/ml streptomycin, and 10% FBS. To assess the effect of CC-I on the ovarian cancer cells, the cells were cultured overnight at a density of 5,000 to 10,000 cells per well in 96 well plates. Then the cells were treated with different concentrations of CC-I (Formula IV) or Formula V for another 3-6 days before cytotoxicity assay using MTS or SRB assay. The IC50 or LD50, 50% inhibition or lethal dose, of compounds is also determined using statistical software (GraphPad) as a general indicator of toxicity.
The IC50 data indicate that CC-I is cytotoxic not only to gliomas but also other types of cancers such as neurofibromas, ovarian cancer, and neuroblastoma. As shown in Table 1, for example, CC-I was toxic to ovarian (OVCAR-3, IC50=3.5/4.3 μM) cancer and breast cancer cells lines.
9
Culturing and Passaging Ovarian Cancer Cells
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We obtained A2780 cells from Dr Paul Modrich (Duke University), the HEY cells from Dr Gloria Huang (Albert Einstein College of Medicine) and SKOV3 cell lines from the American Type Culture Collection, and cultivated them in DMEM media supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin (recommended media). Cells were cultured in a humidified atmosphere (5% CO2) at 37 °C. Trypsin (0.25%)/EDTA solution was used to detach the cells from the culture flask for passaging.
10
Evaluating CC-I Cytotoxicity on Ovarian Cancer
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Example 9
Human ovarian adenocarcinoma SKOV-3 cell lines were ordered from American Type Culture Collection (ATCC, Manassas, Va.), and maintained in McCoy's 5A media with 100 U/ml penicillin, 100 μg/ml streptomycin, and 10% FBS. To assess the effect of CC-I on the ovarian cancer cells, the cells were cultured overnight at a density of 5,000 to 10,000 cells per well in 96 well plates. Then the cells were treated with different concentrations of CC-I (Formula IV) or Formula V for another 3-6 days before cytotoxicity assay using MTS or SRB assay. The IC50 or LD50, 50% inhibition or lethal dose, of compounds is also determined using statistical software (GraphPad) as a general indicator of toxicity.
The IC50 data indicate that CC-I is cytotoxic not only to gliomas but also other types of cancers such as neurofibromas, ovarian cancer, and neuroblastoma. As shown in Table 1, for example, CC-I was toxic to ovarian (OVCAR-3, IC50=3.5/4.3 μM) cancer and breast cancer cells lines.
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