Rat pheochromocytoma 12 (PC12) cells cultured in high glucose-DMEM medium (Hyclone, China) supplemented with 10% fetal bovine serum (TBD science, China) and 1% penicillin-streptomycin (Hyclone), and were placed in an incubator with 5% CO2 and 95% humidity at 37 °C. For evaluating the cytotoxicity, SP-1-3 element leaching liquors were prepared as ISO 10993-12:2012 (biological evaluation of medical devices) after steam sterilization, which were collected by immersing the material in a DMEM medium with a weight-to-volume ratio of 0.2 g ml−1 and incubating them in a cell incubator at 37 °C with 5% CO2 for 72 h before cell counting kit-8 (CCK8) experiment and live/dead cell staining (FDA and PI) on day 1, 3, and 5.
Fetal bovine serum (fbs)
Manufactured by TBD Science
Sourced in China
Fetal bovine serum is a common laboratory reagent used as a cell culture supplement. It is derived from the blood of bovine fetuses and provides a complex mixture of proteins, growth factors, and other nutrients to support the growth and proliferation of a wide range of cell types in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Dmem f 12, by TBD Science (1 mentions)
Mda mb 231, by Thermo Fisher Scientific (1 mentions)
Galunisertib, by Selleck Chemicals (1 mentions)
Tgf β1 recombinant protein, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Mda mb 231, by National Collection of Authenticated Cell Cultures (1 mentions)
Mcf 10a, by National Collection of Authenticated Cell Cultures (1 mentions)
Arpe 19 cells, by Thermo Fisher Scientific (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Arpe 19 cell, by American Type Culture Collection (1 mentions)
Penicillin, by Solarbio (1 mentions)
Streptomycin, by Solarbio (1 mentions)
Rpmi 1640 culture medium, by Thermo Fisher Scientific (1 mentions)
Gsk3β, by Bioworld Technology (1 mentions)
Pgex 4t 1, by GE Healthcare (1 mentions)
Cd31 antibody, by Bioworld Technology (1 mentions)
P mtor, by Bioworld Technology (1 mentions)
β catenin, by Bioworld Technology (1 mentions)
11 protocols using fetal bovine serum (fbs)
1
Cytotoxicity Evaluation of SP-1-3 Leachates
2
Cell Culture Protocols for Cancer Research
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CHO (Chinese hamster ovary cells), MDA-MB-231 (human breast cancer cells), MDA-MB-435S (human breast cancer cells) and MCF-10A (human mammary gland epithelial cells) were obtained from the Chinese Academy of Sciences Shanghai Institute for Biological Sciences-Cell Resource Center, which had characterized the cell lines by short tandem repeat profiling, cell morphology and karyotyping assay. CHO and MDA-MB-231 cells were cultured in Dulbecco's modified Eagle's medium (DMED, Gibco BRL, Rockville, MD, USA), MDA-MB-435S cells were cultured in Leibovitz's L-15 and MDA-MB-231 were cultured in DMEM/F-12 supplemented with 10% fetal bovine serum (TBD Science, Tianjin, China), 100 U/ml penicillin and 100 μg/ml streptomycin (Invitrogen, San Diego, CA, USA), at 37 °C, 5% CO2. The TSP50-stable-expression CHO cell strain was obtained previously.22 (link)
3
Endometrial Cell Culture Under Hypoxia
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The endometrial tissue samples used for the primary culture were removed and transported immediately to the laboratory. They were chopped to a size of 1 mm3 and washed with PBS three times. After the cells were fully digested with trypsin/EDTA (TBD Science, Tianjin, China) in a humidified atmosphere of 5% CO2 at 37°C, the cells were pelleted by centrifugation for 5 min, vigorously resuspended in RPMI 1640 (Gibco; Shanghai, China) supplemented with 10% fetal bovine serum (TBD Science, Tianjin, China) and 100 U/ml penicillin (Gibco, Shanghai, China) and 100 μg/ml streptomycin (Gibco, Shanghai, China), plated, and allowed to settle for up to 3 days. All the cells were identified to be epithelial cells and interstitial cells using immunofluorescence. The endometrial cells were exposed to 1% O2 hypoxia in the presence or absence of 10 ng/ml TGF-β1 recombinant protein (Peprotech, NJ, USA) or 10 μmol/L of the TGF-β1 signal pathway inhibitor galunisertib (LY2157299) (Selleck Chemicals, Houston, USA). Hypoxic conditions were maintained using a modular incubator chamber (Hinasama, Tokyo, Japan) with 5% CO2 and 1% O2 balanced with N2 gas. Data from cells cultured in 21% O2 were used as normal controls.
4
Cell Culture Protocol for Cancer and Normal Cell Lines
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Four cell lines, namely HepG2 (human hepatocellular carcinoma cell line), L02 (human normal liver cell line), MDA-MB-231 (human breast cancer cell line), and MCF-10A (human normal breast cell line), were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in an appropriate medium supplemented with 10 % fetal bovine serum (TBD Science, Tianjin, China), 100U/ml penicillin and 100 mg/mL streptomycin (Ameresco, US) at 37 °C and 5 % CO2.
5
Cultivation of Human NSCLC Cell Lines
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Human NSCLC A549 and H1299 cell lines were obtained from Jilin University. A549 cells were cultured in high glucose DMEM (HyClone, Los Angeles, USA), and H1299 cells were incubated in RPMI-1640 (HyClone, Los Angeles, USA). All culture media were supplemented with 10% fetal bovine serum (tbd Science, Tianjin, China) and 100 units/mL penicillin and streptomycin (HyClone, Los Angeles, USA) and were then cultured in a humidified atmosphere of 5% CO2 at 37°C.
6
A549 Cell Culture Protocol
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Human lung adenocarcinoma cell line (A549) was purchased from ATCC and cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum (TBD Science, Tianjin, China), 1 mmol/L glutamine, 100 U/mL penicillin and10 mg/mL streptomycin (Ameresco, USA), at 37°C, 5% CO2.
7
Optimizing Ox-LDL-Induced ARPE-19 Cell Responses
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ARPE-19 cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). ARPE-19 cells (1.5 × 105 cells/ml) were grown in DMEM/F12 medium (Gibco, New York, NY, USA) supplemented with 10% FBS (TBD science, Tianjin, China), 1% penicillin, and streptomycin (Solarbio) in 37°C with 5% CO2 incubator. The optimal ox-LDL concentration for the following studies was 100 μg/ml, which was chosen based on the mRNA expression levels of NLRP3 and VEGF. The optimal concentration is corresponding to the previous report [25 (link)]. ARPE-19 cells were exposed to 100 μg/ml of ox-LDL for 24 hours with 2-hour pretreatment of 1 μM A740003. Besides, ARPE-19 cells were pretreated with 200 μM of A740003 for 2 hours, followed by stimulation of 100 μg/ml ox-LDL for 48 hours. The optimal concentrations of A740003 for 24/48 hours exposure to ox-LDL were screened by quantitative real-time PCR (Figure S1 ).
8
Culturing Gastric Adenocarcinoma MGC-803 Cells
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Gastric adenocarcinoma cell line MGC-803 was kindly provided by Professor Ke Yang (Beijing University, Health Center, Beijing, China). Cells were maintained in RPMI1640 culture medium (Invitrogen, USA), which was supplemented with 10% heat-inactivated fetal bovine serum (FBS, TBD Science, China), 100U/ml penicillin, and100 U/ml streptomycin, and cultured in a humidified atmosphere of 5% CO2 at 37°C.
9
Investigating ILK-Mediated Signaling Pathways
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EJ cells, HEK 293 cells, pcDNA3.1(−)-myc-RI, pGEX-4 T-RI and pEGFP-C1-RI plasmids were conserved and prepared by our laboratory. pGEX-4 T-1 was bought from GE Healthcare China. pEYFP-N1vector was from Clontech. pCMV-3xflag-CMVTM-10 was purchased from Sigma. BALB/C nude (nu/nu) mice were obtained from Beijing HFK Bioscience Company (Beijing, PR China). FBS was from TBD Science (Tianjin, PR, China). DMEM/High glucose medium, RPMI 1640 medium and G418 were purchased from Gibco-BRL (Carlsbad, CA, USA). Lipofectamine 2000 was bought from Invitrogen, Inc., (Carlsbad, California). Monoclonal mouse antibody of anti-human ILK was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit anti-human β-actin, CD31 antibody, Monoclonal primary rabbit antibody of anti-human PI3K, p-PI3K, PTEN, p- PTEN, Akt, p-Akt, GSK3β, p-GSK3β, mTOR, p-mTOR and β-catenin were obtained from Bioworld Technology, Inc. (St. Louis, USA). The rest of the primary antibodies are from Proteintech Group, Inc (Chicago, IL, USA). Cell Counting Kit-8 was bought from Genview Scientific, Inc (Craigieburn, VIC, AUS).
10
Transient HEK 293 Cell Transfection
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HEK 293 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Biowhittaker) supplemented with 2 mM L-Glutamine (Gibco), 1 mM Sodium Pyruvate (Gibco) and 10% fetal bovine serum (FBS, TBD Science). Cells were transfected one day after plated onto coverslips (Φ18 mm; Deckglasser) and imaged 2 days after transfection.
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