Ear draining lymph nodes from each mouse were carefully and aseptically collected, macerated individually using a cell strainer and the cell suspensions were centrifuged at 400 x g for 10 minutes at 4°C. Cells were counted, adjusted to a concentration of 1x106 cells/well and cultured in complete RPMI [RPMI 1640 supplemented with 10% inactivated fetal bovine serum (Sigma-aldrich), 2 mM/ml-glutamine, 100 µl/ml penicillin, 100 µg/ml streptomycin (Sigma-aldrich)] stimulated or not with live stationary-phase L. braziliensis promastigotes at a ratio 1:5 parasites for 72h at 37°C under 5% CO2. Supernatants were collected and levels of IL-10, IFN-γ, IL-4 and TGF-β were determined by ELISA using a commercial kit (Ebioscience).
Inactivated fetal bovine serum
Manufactured by Merck Group
Sourced in United States
Inactivated fetal bovine serum is a cell culture reagent derived from the blood of fetal bovine. It is a commonly used supplement in cell culture media to provide essential nutrients, growth factors, and other components necessary for the growth and maintenance of cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium f12, by Thermo Fisher Scientific (1 mentions)
Skg iiia, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
3 protocols using inactivated fetal bovine serum
1
Lymph Node Cytokine Profiling in Leishmaniasis
2
Culturing Human Cervical Cancer Cell Lines
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The HPV-16-positive human cervical cancer cell lines, BOKU and SKG-IIIa, and the human immortalized cell line, 293, were purchased from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). The HPV-16-positive human cervical cancer cell line, SiHa, was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). All cell lines were authenticated by the above-mentioned cell banks by means of short-tandem repeat-polymerase chain reaction (pCR) profiling. The cells were cultured in Dulbecco’s modified Eagle’s medium/F12 (life Technologies, Carlsbad, CA, USA) containing 10% inactivated fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin/streptomycin (life Technologies) in 5% carbon dioxide at 37°C.
3
T. cruzi Survival under Blue LED
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Pure cell cultures of T. cruzi (Y and CL strains) were maintained at 25°C in liver infusion tryptose (LIT) broth medium (Becton-Dickinson, NJ, EUA) supplemented with 10% (v/v) inactivated fetal bovine serum (Sigma-Aldrich, San Luis, MS, EUA). These cultures were exposed to blue LED light (460 nm and 40 µW/cm2) for 5 days, 6 h per day, at 24°C. The parasites were counted in 10 µL volume every day for quantification and analysis of their survival using the Neubauer chamber. All the experiments were repeated twice.
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