HCT-116 p53+/+ and HCT-116 p53−/− (ATCC®) cells were grown in Dulbecco’s modified Eagle’s medium (EuroClone, Milan, Italy) supplemented with 10% fetal bovine serum (EuroClone). CH12F3+/+/Δ and CH12F3Δ/Δ/Δ cells were grown in RPMI 1640 (EuroClone) supplemented with 10% fetal bovine serum (EuroClone), 1X non Essential Amino Acids (EuroClone), 1 mM Sodium Pyruvate (EuroClone), 25 mM HEPES (EuroClone) and 50 μM β-mercaptoethanol (Promega, Madison, WI, USA). OCI-AML2 cells were grown in α-MEM (Invitrogen, Carlsbad, CA, USA) supplemented with 20% fetal bovine serum (EuroClone). HCC70 cells were grown in RPMI 1640 (EuroClone) supplemented with 10% fetal bovine serum (EuroClone). All culturing media were also supplemented with 2 mM GlutaMAX (EuroClone), 100 U/ml penicillin and 10 μg/ml streptomycin (EuroClone).
Fetal bovine serum (fbs)
Manufactured by Euroclone
Sourced in Italy, United States, United Kingdom, Switzerland, Germany, Belgium, France
Fetal Bovine Serum (FBS) is a widely used cell culture supplement derived from the blood of bovine fetuses. It provides a complex mix of proteins, growth factors, and other essential nutrients necessary for the growth and maintenance of various cell types in vitro.
Automatically generated - may contain errors
Lab products found in correlation
L glutamine, by Euroclone (164 mentions)
Streptomycin, by Euroclone (117 mentions)
Penicillin, by Euroclone (116 mentions)
Penicillin streptomycin, by Euroclone (115 mentions)
Dulbecco modified eagle medium (dmem), by Euroclone (90 mentions)
Glutamine, by Euroclone (53 mentions)
Rpmi 1640 medium, by Euroclone (51 mentions)
Rpmi 1640, by Euroclone (49 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (46 mentions)
L glutamine, by Merck Group (45 mentions)
L glutamine, by Lonza (38 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (38 mentions)
Penicillin, by Thermo Fisher Scientific (35 mentions)
Streptomycin, by Merck Group (34 mentions)
Streptomycin, by Thermo Fisher Scientific (34 mentions)
Penicillin, by Merck Group (33 mentions)
Penicillin streptomycin, by Lonza (30 mentions)
Penicillin streptomycin, by Merck Group (28 mentions)
L glutamine, by Thermo Fisher Scientific (27 mentions)
Dmem high glucose, by Euroclone (27 mentions)
851 protocols using fetal bovine serum (fbs)
1
Cell Culture Conditions for Cancer Cell Lines
2
Culturing Human Melanoma and Endothelial Cells
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Human metastatic melanoma cell line A375 (ATCC, Manassas, VA, USA) was grown in high D-glucose DMEM, with 10% (v/v) heat-inactivated fetal bovine serum (Euroclone Milan, Italy), 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mmol/L glutamine in a humidified atmosphere with 5% CO2 in air. The culture medium was changed every 2 days. HUVECs (ATCC, Manassas, VA, USA) were grown in Endothelial Basal medium, supplemented with 10% FBS (Euroclone, Milan, Italy), on gelatin-coated dishes. Endothelial Colony Forming Cells (ECFCs) were isolated from >50 mL human umbilical cord blood (UCB) of healthy newborns after maternal informed consent as previously described [20 (link),46 (link)] The purification and use of stem cells from cord blood for research purposes is permitted by an Italian law after obtaining informed consent from the mothers (art. 2, paragraph 1, letter f, decree of 18 November 2009). ECFCs were grown in EGM-2 culture medium (Lonza, Lonn, Swiss), supplemented with 10% FBS (Euroclone, Milan, Italy) onto gelatin-coated dishes. ECFCs were grown in a humified atmosphere with 5% of CO2 in air and the medium was refreshed every 2 days.
3
Ovarian Cancer Cell Line RNA Interference
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The human ovarian carcinoma cell line SKOV-3 was obtained from the CEINGE Cell Culture Facility (Naples, Italy) and was grown in RPMI-1640 medium (Euroclone) containing 10% fetal bovine serum (Euroclone). The immortalized Fallopian tube secretory epithelial cell line FT194 was kindly provided by Dr. R. Drapkin (Boston, USA) and was maintained in DME-F12 medium (Euroclone) containing 2% Ultroser G serum (PALL). The OVCAR-3 cell line was obtained from ATCC and was maintained in RPMI-1640 medium (Euroclone) containing 20% fetal bovine serum (Euroclone) and 0.01 mg/ml bovine insulin. PEA1 and PEO14 cells were purchased from Sigma-Aldrich and mantained in RPMI-1640 medium (Euroclone) containing 10% fetal bovine serum (Euroclone), 2 mM glutamine and 2 mM sodium pyruvate (Gibco, Life Technologies).
For RNA interference, SKOV-3, FT194 cells and PEA1 were plated at 2 × 105 cells/60-mm tissue culture dish 24 h prior to transfection and were transfected in replicates with 5 nM PAX8 siRNA (Ambion, Life Techonologies, siRNA ID s15403) or siRNANon-Targeting (Ambion, Life Technologies, siRNA ID 4390843) as scramble, using the Lipofectamine RNAiMAX transfection reagent (Invitrogen) following the manufacturer's protocol. Cells were harvested 24 h after transfection and the total RNA was prepared. Human Fallopian tubes RNA was from Origene (CR559726).
For RNA interference, SKOV-3, FT194 cells and PEA1 were plated at 2 × 105 cells/60-mm tissue culture dish 24 h prior to transfection and were transfected in replicates with 5 nM PAX8 siRNA (Ambion, Life Techonologies, siRNA ID s15403) or siRNANon-Targeting (Ambion, Life Technologies, siRNA ID 4390843) as scramble, using the Lipofectamine RNAiMAX transfection reagent (Invitrogen) following the manufacturer's protocol. Cells were harvested 24 h after transfection and the total RNA was prepared. Human Fallopian tubes RNA was from Origene (CR559726).
4
Culturing Cell Lines for Research
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The parental MCF-7 (breast cancer), MDA-MB-231 (triple-negative breast cancer), U-87 MG (glioblastoma), A549 (non-small cell lung cancer), and PANC-1 (pancreatic cancer) cell lines and HDFs (human dermal fibroblasts) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). MCF-7 and A549 cells were cultured as an attached monolayer and maintained in RPMI 1640 medium (EuroClone, Milan, Italy) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS) (EuroClone, Milan, Italy), 1% penicillin-streptomycin (EuroClone, Milan, Italy), and 2 mM L-glutamine. MDA-MB-231 cells were cultured as an attached monolayer and maintained in MEM (EuroClone, Boston, MA, USA) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS) (EuroClone, Milan, Italy), 1% penicillin-streptomycin (EuroClone, Milan, Italy), and 2 mM L-glutamine. U-87, PANC-1 and HDFs were cultured as an attached monolayer and maintained in DMEM (EuroClone, Milan, Italy) supplemented with 10% (v/v) heat-inactivated FBS (EuroClone, Milan, Italy), 1% penicillin-streptomycin (EuroClone, Milan, Italy), and 2 mM L-glutamine. All cells were incubated at 37 °C in a 5% CO2 tissue culture incubator (MEMmert, Schwabach, Germany).
5
Pancreatic Cancer Cell Culture Conditions
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Cells were cultured at 37 °C with 5% CO2 in a humidified atmosphere. PANC-1 were cultured in DMEM (Euroclone, Milan, Italy) supplemented with 4 mM of L-glutamine and 10% FBS (Euroclone). MiaPaca-2 and BxPc-3 were cultured in RPMI (Euroclone) supplemented with 2 mM of L-glutamine and 10% FBS (Euroclone). PSC-RLT were cultured in DMEM F-12 (Euroclone) supplemented with 2 mM of L-glutamine and 10% FBS (Euroclone). HPDE were cultured in 50% RPMI 1640 (Life Technologies, Carlsbad CA, USA), 50% Keratinocyte medium—SFM (Life Technologies) supplemented with FBS 10% heat inactivated, MEM Non-Essential Amino Acids 1 × (Life Technologies), Pen/Strep 1X, Hepes 10 mM (Life Technologies), bovine pituitary extract 0.025% (Life Technologies), and EGF human recombinant 2.5 ug/L (Life Technologies).
PANC-1, MiaPaca-2 and BxPc-3 cells were obtained from the American Type Culture Collection (ATCC); HPDE were kindly gifted by Prof. I. Szabò (University of Padua, Italy); PSC-RLT cells were kindly gifted by Prof. F. Alves (UMG, Department of Hematology and Medical Oncology and the Institute for Diagnostic and Interventional Radiology, Goettingen, Germany). When cultured as spheroids, cells were seeded on an agarose base layer (1.5 g/L) in 96 wells plates and grown for 72 h.
PANC-1, MiaPaca-2 and BxPc-3 cells were obtained from the American Type Culture Collection (ATCC); HPDE were kindly gifted by Prof. I. Szabò (University of Padua, Italy); PSC-RLT cells were kindly gifted by Prof. F. Alves (UMG, Department of Hematology and Medical Oncology and the Institute for Diagnostic and Interventional Radiology, Goettingen, Germany). When cultured as spheroids, cells were seeded on an agarose base layer (1.5 g/L) in 96 wells plates and grown for 72 h.
6
Breast, Lung, and Pancreatic Cancer Cell Lines
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Human breast adenocarcinoma SKBR3 cells (p53 mutated in R175H) were maintained in DMEM medium containing 10% fetal bovine serum (FBS; EuroClone, Milan, Italy), human lung adenocarcinoma H1975 cells (p53 R273H) were cultured in RPMI 1640 medium containing 10% FBS (EuroClone, Milan, Italy), and human pancreatic carcinoma MIA PaCa 2 cells (p53 R248W) were maintained in DMEM medium supplemented with 10% FBS (EuroClone, Milan, Italy), 2.5% horse serum (Thermo Fisher Scientific, Waltham, MA, USA), and 1 mM of sodium pyruvate (Sigma-Aldrich, St. Louis, MO, USA). The cell lines described above were purchased from ATCC. Treatments with etoposide (Sigma-Aldrich) were performed on SKBR3, MIA PaCa 2, and H1975 at 25 μM for the indicated time points. Lipofectamine® LTX (Thermo Fisher Scientific, Waltham, MA, USA) was used to transfect the SKBR3, MIA PaCa 2, and H1975 in accordance with the manufacturer’s instructions.
7
Culturing Human Lung Cell Lines and Propagating 229E Coronavirus
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Low-passage human embryonic lung fibroblasts (HELFs) were prepared and grown as monolayers in MEM supplemented with 10% FBS (Euroclone), 1 mM sodium pyruvate, 2 mM glutamine, 100 U mL−1 penicillin, and 100 μg/mL 1 streptomycin sulfate, as previously described [58 (link)]. MRC5 (ATCC® CCL-171™) lung fibroblast were purchased from the American Type Culture Collection (ATCC) and cultured in Eagle’s minimum essential medium (MEM; EuroClone) supplemented with 10% fetal bovine serum (FBS, Euroclone), 2 mM glutamine, 1 mM sodium pyruvate, 100 U/mL penicillin, and 100 µg/mL streptomycin sulfate (P/S, both from Euroclone). Lung epithelial A549 (ATCC® CCL-185™) cells were purchased from ATCC, and cultured in Dulbecco’s modified Eagle medium (DMEM; EuroClone), supplemented with 10% fetal bovine serum (FBS, Euroclone), 2 mM glutamine, 1 mM sodium pyruvate, 100 U/mL penicillin, and 100 µg/mL streptomycin sulfate (P/S, both from Euroclone). The human coronavirus 229E (ATCC® VR-740™) was purchased from the ATCC and propagated and titrated on MRC5 cells.
8
Cytotoxicity Evaluation of Natural Compounds
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Acetonitrile for the HPLC-DAD-MS analysis was purchased by Merck (Darmstadt, Germany), while water was purified with a Direct-Q UV system by Millipore (Darmstadt, Germany). Chlorogenic acid, rutin and oleuropein (analytical purity) were purchased from Sigma Aldrich (Darmstadt, Germany). All other chemicals used were purchased from Alfa-Aesar, Karlsruhe, Germany. The cytotoxicity was tested on two tumoral cell lines: murine melanoma cells (B16F10 cells) and human cancer cell line (HeLa). The selected cell lines were obtained from American Type Cell Collection (ATCC) (Manassas, VA, USA). The B16F10 cells were maintained in RPMI 1640 medium (Sigma Aldrich, Darmstadt, Germany) supplemented with 10% fetal bovine serum (EuroClone, MI, Pero, Italy) and 1% Antibiotic-Antimycotic 100× (Sigma Aldrich, Darmstadt, Germany). Hela cells were maintained in DMEM/F-12 medium (Gibco Life Technologies, Paisley, UK) supplemented with 10% fetal bovine serum (EuroClone, MI, Pero, Italy) and 1% Antibiotic-Antimycotic 100× (Sigma Aldrich, Darmstadt, Germany). The two cell lines were cultured in a 5% CO2 incubator (Advantage-Lab, Schilde, Belgium) at 37 °C in a humidified atmosphere. Cisplatin (Ebewe Pharma Ges.m.b. H. Nfg. KG, Unterach am Attersee, Austria) was included as standard positive control for cytotoxicity assay.
9
Cell Culture Conditions for Hematological Malignancies
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OCI/AML2 and 3 cells were grown in alpha-MEM (Euroclone, Milan, Italy) supplemented with 20% fetal bovine serum, 100 U/mL penicillin and 10 µg/mL streptomycin sulfate. Jurkat and HL-60 cells were grown in RPMI-1640 medium (Euroclone, Milan, Italy); all cells were supplemented with 10% fetal bovine serum (FBS, EuroClone, Milan, Italy), L-glutamine (2 mM) (Euroclone, Milan, Italy), penicillin (100 U/mL) and streptomycin (100 mg/mL) (Euroclone, Milan, Italy), and were cultured in a humidified incubator, at 37 °C, in a 5% v/v CO2 atmosphere. All chemical reagents used for treatments were supplied from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise specified.
10
Immortalized Hepatic Stellate Cell Lines
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LX2 (a human immortalized hepatic stellate cell line), HSC-T6 (a rat immortalized hepatic stellate cell line), and HeLa (a human cervical carcinoma cell line) cells were cultured at 37°C in an atmosphere of 5% CO2 in Dulbecco's Modified Eagle Medium (Euroclone - Milan, Italy) supplemented with 10% Fetal Bovine Serum (Euroclone), 2 mM L-Glutamine and antibiotics. HepG2 cells (a human hepatocarcinoma cell line) were cultured at 37°C in an atmosphere of 5% CO2 in Eagle's Minimum Essential Medium (Euroclone) containing 10% Fetal Bovine Serum, 2 mM L-Glutamine and antibiotics.
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