Skov-3, HK2 and NIH-3T3 cells were purchased from the Shanghai Cell Bank of Type Culture Collection at the Chinese Academy of Sciences. Skov-3 cells (104 cells/well) were seeded in 96-well flat bottom plates and maintained in 1640 complete medium, and cultured overnight at 37 °C and 5% carbon dioxide. HK2 and NIH-3T3 were cultured in DMEM complete medium, and their culture environment were the same as Skov-3. Cell viability was evaluated by CCK-8 kits or Calcein AM/PI Detection kits.
Skov3
Manufactured by Shanghai Cell Bank
Sourced in China
SKOV3 is a human ovarian adenocarcinoma cell line. It is a commonly used in vitro model for ovarian cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Ho8910, by Shanghai Cell Bank (5 mentions)
Ovcar 3, by Shanghai Cell Bank (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Mcf 7, by Shanghai Cell Bank (2 mentions)
Caov3, by Shanghai Cell Bank (2 mentions)
A2780, by Shanghai Cell Bank (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Iose80, by Shanghai Cell Bank (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Mccoy s 5a medium, by Merck Group (1 mentions)
Carboplatin, by Qilu Pharmaceutical (1 mentions)
Lipofectamine 2000 lipo 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Branched pei, by Merck Group (1 mentions)
Bicinchoninic acid bca assay kit, by Beyotime (1 mentions)
Triethylamine, by Merck Group (1 mentions)
Hct116, by Shanghai Cell Bank (1 mentions)
Hepg2, by Shanghai Cell Bank (1 mentions)
Hela cells, by Shanghai Cell Bank (1 mentions)
19 protocols using skov3
1
Cell Viability Assay Protocol
2
Cell Culture and Hypoxia Conditions
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SKOV3 and HO8910 cell lines were purchased from the Shanghai Cell Bank of Chinese Academy of Sciences. The cells were cultured in McCoy’s 5A medium (Sigma, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Australia), 100 lg/ml streptomycin and 100 units/ml penicillin. Cultures were maintained at 37 °C in a humidified atmosphere of 5% CO2. The oxygen concentration of hypoxia condition was 1% O2.
3
Evaluating SKOV3 Cell Cytotoxicity
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The human serous cystadenocarcinoma ovarian cancer cell line SKOV3 (BNCC310551) was purchased from the Shanghai cell bank (Shanghai, China). MTT cytotoxic kit was purchased from Wuhan BOSTER Biological Technology Co., LTD (Wuhan, Hubei Province, China). Indoleamine 2,3 dioxygenase kit was purchased from the Elabscience Biotechnology Co., LTD (Wuhan, Hubei Province, China). Carboplatin was purchased from Qilu pharmaceutical Co., LTD (Jinan, Shandong Province, China). Matrigel matrix adhesive was purchased from BD company of America (Franklin Lake, New Jersey, USA). Lactate dehydrogenase (LDH) assay kit was purchased from Nanjing Bioengineering Institute (Nanjing, Jiangsu Province, China). The CD8+ T cell separation kit was purchased from STEMCELL Company (Beijing, China) and the ELISPOT kit of CD8+ T cells was purchased from RD Company (Minnesota, USA). Human peripheral blood was collected from the group of experimental healthy volunteers.
4
Branched PEI and PDLLA Nanocarrier for siRNA Delivery
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Branched PEI (Mw ~25 kDa, average Mn ~10 kDa) and poly(D,L-lactic acid) (Mw ~15 kDa) were obtained from Sigma-Aldrich and Polysciences Inc. (Germany), respectively. Acryloyl chloride was purchased from TCI (Shanghai) Development Co., Ltd. (Shanghai, China). Triethylamine was provided by Sigma-Aldrich. siRNA targeting PKM2 (siPKM2): 5′-CCAUAAUCGUCCUCACCAA-3′ (sense), and negative control siRNA (siNC) were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China). Cy5-labled siRNA targeting PKM2 was provided by GenePharma Co. (Shanghai, China). Bicinchoninic acid (BCA) assay kit was obtained from Beyotime Institute of Biotechnology (Shanghai, China). Lipofectamine® 2000 (Lipo 2000) Reagent was provided by Invitrogen Life Technologies (Carlsbad, CA, USA). Primary antibodies against PKM2 was purchased from Cell Signaling Technology (Danvers, MA, USA). All other organic solvents used were of analytical grade. HCT116, HepG2 and SKOV3 cells were obtained from Shanghai Cell Bank of Chinese Academy of Sciences (Shanghai, China).
5
Cell Line Cultivation Protocol
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The following cells lines were purchased
from the Shanghai Cell Bank of the Chinese Academy of Sciences: HEK
293T cells (human embryonic kidney transformed cells), MCF 10A cells
(human normal mammary epithelial cells), MCF-7 cells (human breast
cancer cell), MCF-7 (PDX) cells (human breast cancer cell), SKOV3
(human ovarian cancer cell), and HeLa cells (human cervical cancer
cells). Live cells were kept at 37 °C in a humidified atmosphere
containing 5% CO2 and cultivated in Dulbecco’s modified
Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS), 1%
penicillin, and 1% streptomycin sulfate. MCF 10A cells were cultivated
in DMEM/F12 containing 5% horse serum, 20 ng/mL EGF, 0.5 μg/mL
hydrocortisone, 10 μg/mL insulin, 1% NEAA, and 1% P/S.
from the Shanghai Cell Bank of the Chinese Academy of Sciences: HEK
293T cells (human embryonic kidney transformed cells), MCF 10A cells
(human normal mammary epithelial cells), MCF-7 cells (human breast
cancer cell), MCF-7 (PDX) cells (human breast cancer cell), SKOV3
(human ovarian cancer cell), and HeLa cells (human cervical cancer
cells). Live cells were kept at 37 °C in a humidified atmosphere
containing 5% CO2 and cultivated in Dulbecco’s modified
Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS), 1%
penicillin, and 1% streptomycin sulfate. MCF 10A cells were cultivated
in DMEM/F12 containing 5% horse serum, 20 ng/mL EGF, 0.5 μg/mL
hydrocortisone, 10 μg/mL insulin, 1% NEAA, and 1% P/S.
6
Culturing Human Ovarian Cancer Cell Lines
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Five human cell lines containing OC (C200, Caov3, SKOV3, HO8910, and OVCAR3) were obtained from Shanghai Cell Bank, Chinese Academy of Science (Shanghai, China). All cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Hyclone, USA), supplemented with 100 μg/ml streptomycin, 10% heat-inactivated FBS, and 100 U/ml penicillin. All media were incubated in an incubator (Thermo Fisher Scientific Inc., Waltham, MA, USA) (37°C, 100% humidity, and 5% CO2).
7
In Vitro Culture of Ovarian Cancer Cell Lines
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The human ovarian cancer cell lines A2780 and SKOV3 were purchased from the Shanghai Cell Bank of Chinese Academy of Sciences (Shanghai, China). Both cells lines were cultured in RPMI 1640 (Gibco Life Technologies, Carlsbad, CA, USA),supplemented with 10% fetal bovine serum(Invitrogen, Carlsbad, CA, USA), 100 IU/mL streptomycin and 100 IU/mL penicillin, at 37 °C under humidified air containing 5% CO2.
8
Artesunate and Cisplatin Effects on Ovarian Cancer Cells
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Human ovarian cancer cell lines HO8910 and SK-OV-3 were from Shanghai Cell Bank, Chinese Academy of Sciences. Human ovarian carcinoma cell line A2780 was obtained from Sigma-Aldrich. Immortalized fallopian epithelial cell line FTE-187 was as described.23 Normal human fibroblasts (NHF) were generated in this lab.24 A2780 and HO8910 cells were cultured in RPMI 1640, NHF in DMEM, and FTE-187 in M199, supplemented with 10% fetal bovine serum, 100 units/ml penicillin, and 100 mg/ml streptomycin. All the cells were incubated in a humidified atmosphere of 95% air and 5% CO2. Artesunate was acquired from Sigma-Aldrichand was dissolved in dimethyl sulfoxide (DMSO)(25 mg/ml) as a stock solution and stored at −20°C in aliquots until use. It was applied to the cultured cells at the concentration of 0, 5, 10, 25, or 50 µg/ml for various periods. Cisplatin, also purchased from Sigma-Aldrich, was dissolved in PBS.
9
Establishment of Siva 1 Overexpressing Ovarian Cancer Cells
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Ovarian cancer SKOV3 and OVCAR-3 cell lines were purchased from the Shanghai Cell Bank of Chinese Academy of Sciences and cultured with RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA USA) supplemented with 10% fetal bone serum (FBS; Hyclone; GE Healthcare; Logan, UT, USA) at 37°C in 5% CO2. A2780 cells were purchased from Cell Preservation Center of Wuhan University and cultured with Dulbecco's modified Eagle's medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented 10% FBS at 37°C in 5% CO2. Siva 1 overexpression plasmid pCMV3-Siva 1 or the control pCMV3 [China National Pharmaceutical Group (CNPGC); Sinopharm, Beijing, China] were transfected into A2780 cells with lipofectamine reagent (Invitrogen; Thermo Fisher Scientific, Inc.). A total of 24 h after transfection, 200 µg/ml G418 (Invitrogen; Thermo Fisher Scientific, Inc.) was added to the medium, and the culture medium was renewed every 2 days. Approximately 1 week later, monoclonal cell clusters were observed and selected for culture. Thereafter, the Siva 1 stably expressed A2780 cell line and its control were used for subsequent experiments.
10
Culturing Ovarian Cancer Cell Lines
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HEK293, SKOV3, ES-2 and HO8910 cells were purchased from Shanghai Cell Bank of the Chinese Academy of Science (Shanghai, China). SKOV-3 and ES-2 cells were grown in McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS). HEK293 and HO8910 cells were grown in Dulbecco’s modified Eagle medium (DMEM) - high glucose medium (Life, U.S.) supplemented with 10% FBS (Life, U.S.). Ovarian cancer cell lines (OVCAR3) and Anglne cells were purchased from China Center for Type Culture Collection (Wuhan, China). OVCAR3 cells and Anglne cells were respectively grown in DMEM-high glucose medium with 10% FBS and Eagle’s minimal essential medium (Eagle’s MEM) with 10% FBS. A2780 cells were purchased from Nanjing KeyGen Biotech (Nanjing, China) and cultured in a DMEM-high glucose medium with 10% FBS. Primary normal ovarian surface epithelial (NOSE) cells were established through the previously reported methods [15 (link)]. All cell lines were maintained in 5% CO2 at 37 °C.
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