Bovine fetal serum

Manufactured by Merck Group

Sourced in United States

Bovine fetal serum is a cell culture supplement derived from the blood of bovine fetuses. It provides a complex mixture of proteins, growth factors, and other nutrients essential for the growth and proliferation of cells in vitro.

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FBS (13055 citations) Bovine serum (80 citations) FBS (fetal bovine serum) (24 citations) Fetal bovine serum (FCS) (7 citations) Fetal bovine serum superior (7 citations) Non-inactivated fetal bovine serum (4 citations) Inactivated fetal bovine serum (3 citations) Fetal bovine calf serum (2 citations) Fetal bovine serum solution (FBS; (2 citations) Fetal bovine serum (FBS); L-glutamine (L-Glut); (2 citations)

14 protocols using bovine fetal serum

Leishmania infantum Infection Model

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Experiments were performed with the L. infantum strain (MHOM/PT/88/IMT151). Promastigotes were cultured at 26°C until the stationary phase (fourth day) in Schneider’s medium supplemented [L-glutamine (1mM/mL), antibiotic (penicillin: 200μg/mL, streptomycin: 200μg/mL), and bovine fetal serum (10%)] (Sigma Chemical Co., St. Louis, USA).
Hamsters (n = 42) were infected with 2 x 10⁷ promastigotes of L. infantum at the stationary growth phase by intraperitoneal (i.p.) route, in a final volume of 50μL. Uninfected hamsters (n = 30) were used as a control group. The infection was monitored for six months, and the animals were euthanized at 30, 120, and 180 days post-infection (dpi), using a combination of anesthetics. Blood was collected by cardiac puncture, between 6:00 and 8:00 a.m. (nadir of the endogenous circadian rhythm) [21 (link)]. The blood obtained from each animal was split into three samples: 1) a tube containing heparin for plasma obtention; samples were stored at -20°C for cortisol dosage; 2) a tube containing EDTA for measurement of hematological parameters; 3) a clot activator tube to obtaining serum for biochemical analysis (BD Vacutainer, USA).

Barros-Gonçalves T.D., Saavedra A.F., da Silva-Couto L., Ribeiro-Romão R.P., Bezerra-Paiva M., Gomes-Silva A., Carvalho V.F., Da-Cruz A.M, & Pinto E.F. (2021). Increased levels of cortisol are associated with the severity of experimental visceral leishmaniasis in a Leishmania (L.) infantum-hamster model. PLoS Neglected Tropical Diseases, 15(11), e0009987.

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Silica Cytotoxicity in Cell Cultures

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In cell cultures treated with silica, several tests were performed to find the concentration of fluxed silica. To ensure at least 80% viability, silica was insufflated using the flow given by no air. The established flow is 40 µg/hour. In the case of silica and ozone, silica was insufflated using the flow of the ozonator set at an ozone concentration of 120 ppb.
Hs27 (human skin fibroblasts, ATCC CRL-1634) and A549 (lung alveolar adenocarcinoma epithelial, ATCC CCL-185) cell lines were purchased from the American Type Culture Collection.
The cells were seeded in monolayer in Dulbecco’s modified Eagle medium (DMEM) with 0.1 mg/mL streptomycin and 100 UI/mL penicillin, 10% bovine fetal serum and 2 mM L-glutamine (SIGMA, Milan, Italy) at a controlled atmosphere with 5% CO₂, 90% humidity and at 37 °C. The cells were plated to the density of 2500 cells/well for MTS viability test and 10,000 cells/cm² for the micronuclei and comet tests. The cells were detached with 0.05% trypsin–0.02% EDTA. All materials were purchased from SIGMA-Aldrich, Merk Life Science S.r.L., Milan, Italy.

Colafarina S., Di Carlo P., Zarivi O., Aloisi M., Di Serafino A., Aruffo E., Arrizza L., Limongi T, & Poma A. (2022). Genotoxicity Response of Fibroblast Cells and Human Epithelial Adenocarcinoma In Vitro Model Exposed to Bare and Ozone-Treated Silica Microparticles. Cells, 11(2), 226.

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Inflammatory Cytokine Assessment in Intestinal Explants

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Ileum and colon explants were cultured in RPMI 1640 supplemented with 10% bovine fetal serum and antibiotics, in presence or absence of 10 µg/mL of LPS from E. coli as proinflammatory stimulus (all from Sigma Chemical Co., St. Louis, MO, USA) for 24 h at 37°C in an atmosphere of 95% air and 5% CO₂ as described previously (40 (link), 42 (link)). Supernatants were collected, centrifuged, and frozen until cytokine (IL-6, IL-4, IL-10, IL-17A, IFN-γ, and GM-CSF) measurements (Ready-SET-Go!^® ELISA Kit, eBioscience, France). All assays were performed according to the manufacturer’s instructions.

Carasi P., Racedo S.M., Jacquot C., Elie A.M., Serradell M.D, & Urdaci M.C. (2017). Enterococcus durans EP1 a Promising Anti-inflammatory Probiotic Able to Stimulate sIgA and to Increase Faecalibacterium prausnitzii Abundance. Frontiers in Immunology, 8, 88.

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Antipsychotic Effects on Telomere Length

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The PBMCs were maintained in a 6-well dish, in RPMI 1640 medium supplemented with 10% bovine fetal serum (Sigma) and antibiotics (penicillin/streptomycin 1% v/v) at 37°C and an atmosphere with 5% of CO₂. To cause initial telomeric damage, we tested different hydrogen peroxide (H₂ O₂) (Sigma) concentrations: 100, 200, 400, and 600 μM. The cultures with H₂O₂ were maintained for 3 days in the same conditions described.
For each individual culture of PBMCs, we induced oxidative stress with 200 μM of H₂O₂ for 3 days, after which the distinct antipsychotics were added to the culture medium for additional 4 days. We used working concentrations of antipsychotic drugs within the range of plasma therapeutic levels in three experimental (intervention) groups: (a) haloperidol 7 ng/mL (Van Putten et al., 1992); (b) clozapine 1000 ng/mL (Ulrich et al., 2003); (c) aripiprazole 125 ng/mL (McEvoy et al., 2014). Two comparison groups [controls and vehicle (DMSO ≤ 0.02%)] were established: (d) PBMCs maintained for 7 days in culture media only (no H₂O₂ and no antipsychotics), presumably accounting for baseline in vitro effect (no/minimal undisturbed change in telomere length); and (e) PBMCs treated with H₂O₂ for 3 days but no antipsychotics added between days 4 and 7 (negative control), indicative of the effect of oxidative stress on telomeres).

Polho G.B., Cardillo G.M., Kerr D.S., Chile T., Gattaz W.F., Forlenza O.V., Brentani H.P, & De-Paula V.J. (2021). Antipsychotics preserve telomere length in peripheral blood mononuclear cells after acute oxidative stress injury. Neural Regeneration Research, 17(5), 1156-1160.

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Synthesis and Characterization of PLA-PVA Hydrogel

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Polylactic acid (PLA), Polyvinyl alcohol (PVA) Phosphate buffered saline (PBS), PBS/EDTA (10 nM) freshly prepared solution, Freund's complete adjuvant, Histopaque reagent, Pluronic F127, DMEM high glucose, bovine fetal serum, doxorubicin, poly-d-lysine, glucose, HEPES, calcium and magnesium were purchased from Sigma Aldrich (St. Louis, MO, USA). Hydroxychloroquine was purchased from Infinity pharma and azithromycin was purchased from Eurofarma.

de Barros A.O., Pinto S.R., dos Reis S.R., Ricci-Junior E., Alencar L.M., Bellei N.C., Janini L.R., Maricato J.T., Rosa D.S, & Santos-Oliveira R. (2022). Polymeric nanoparticles and nanomicelles of hydroxychloroquine co-loaded with azithromycin potentiate anti-SARS-CoV-2 effect. Journal of Nanostructure in Chemistry, 13(2), 263-281.

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Thyroid Gland Cryopreservation Protocol

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After removal, the thyroid gland was kept in a 1 × sterile PBS solution (pH 7.4). In a laminar flow chamber, a nutrient solution containing 43% RPMI 1640 (Cultilab, Campinas, SP, BR), 50% bovine fetal serum (Sigma, St Louis, Mo, USA), and 7% DMSO (dimethylsulfoxide) was packed in a cryotube previously cooled in ice, as mentioned by Taylor and cols. (2019).²⁷ (link) The cryotube was then placed in a freezer (−4 °C) for 1 hour. At the end of the process, the sample was identified and stored for seven days in liquid nitrogen at −196 °C. To thaw the samples the group used a 37 °C water bath until the sample reached room temperature and two washes were performed with sterile PBS.

Schanaider A., Barboza T., Vasconcellos M., Gutfilen-Schlesinger G, & de Souza S.A. (2022). Rat thyroid graft transplantation after cryopreservation with scintigraphic standardization for an experimental study. Clinics, 77, 100065.

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Osteoclast Formation and Bone Resorption

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Phosphate buffered saline (PBS), PBS/EDTA, bovine serum albumin (BSA), methylated bovine serum albumin (mBSA), Freund’s complete adjuvant, Histopaque reagent, Pluronic F127, TRAcP staining kit, DMEM high glucose, Bovine fetal serum, M-CSF, RANK-L, Doxorubicin, Poly-D-lysine, Glucose, HEPES, Calcium, and Magnesium were purchased from Sigma Aldrich (St. Louis, MO, USA). MayGrünwald and Giemsa dyes were purchased from Merck (Germany). 3% sodium pentabarbital (Hypnol^®) was purchased from Syntec (Brazil). Tevametho (injectable metrotrexate 25 mg/mL) was purchased from Teva Farmacêutica Ltda (Brazil), and Hydroxychloroquine was purchased from Must Check. The radium dichloride (Xofigo^®) was purchased from Bayer.

Yang Y., Alencar L.M., Pijeira M.S., Batista B.D., França A.R., Rates E.R., Lima R.C., Gemini-Piperni S, & Santos-Oliveira R. (2022). [223Ra] RaCl2 nanomicelles showed potent effect against osteosarcoma: targeted alpha therapy in the nanotechnology era. Drug Delivery, 29(1), 186-191.

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Bovine PBMC Isolation and Stimulation

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PBMCs were isolated from heparinized bovine blood by density gradient using Ficoll-Paque ^TM PLUS (GE Healthcare, Uppsala, Sweden). Once purified, PBMCs were seeded in RPMI 1640 culture medium with 2 mM L-glutamine, 25 mM HEPES, 5 × 10⁻⁵ M β-mercaptoethanol, 100 U/mL of penicillin, 100 μg/mL of streptomycin, 2 g/L of sodium bicarbonate, and 10% bovine fetal serum (Sigma-Aldrich, St. Louis, MI, USA, EE. UU.). PBMCs were seeded into 24-well plate at a concentration of 1 × 10⁶ cells/mL and stimulated with 0.3 µg/µL CFPE. Untreated cells (control) were included. Then, PBMCs were cultivated a 37 °C in a 5% CO₂ atmosphere for three and nine days.

Manzo-Sandoval A., Jaramillo-Meza L., Olguín-Alor R., Sánchez-Torres L.E, & Díaz-Otero F. (2023). Application of Multiparametric Flow Cytometry Panels to Study Lymphocyte Subpopulations in Tuberculin-Positive Cattle. Veterinary Sciences, 10(3), 197.

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Evaluating Ursolic Acid and its Derivatives on Human Blood Cell Cultures

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Blood samples from non-smoker healthy donor (B Rh+ blood type) were collected into heparin tubes and used throughout the experiment. The heparinized blood (1.0 mL) in 6.0 mL of Dulbecco’s phosphate buffered saline with MgCl₂ and CaCl₂ medium (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany) supplemented with 10% bovine fetal serum (Sigma-Aldrich, Chemie GmbH, Taufkirchen, Germany), 1% L-glutamine (Merck, KGaA, Darmstadt, Germany), and antibiotics mix solution (100 µL/mL, 10,000 U penicillin, 10 mg streptomycin, 25 µg amphotericin B per 1 mL, Sigma-Aldrich, Chemie GmbH, Taufkirchen, Germany) added in 6 wells untreated Nuncleon plates were incubated in a 37 °C incubator with 5% CO₂. After 72 h of incubation, blood cell cultures were treated with UA, UBA and UBE dissolved in 1% DMSO. Human blood samples were treated with final concentrations of 25, 50, 75, and 125 µg/mL of UA. Higher concentrations (75, 125, 250, and 500 µg/mL) of both UBA and UBE were used to treat the human blood cell cultures. In addition, the blood cells were treated with 1% DMSO as the negative control (solvent control).

Popovici V., Matei E., Cozaru G.C., Aschie M., Bucur L., Rambu D., Costache T., Cucolea I.E., Vochita G., Gherghel D., Caraiane A, & Badea V. (2021). Usnic Acid and Usnea barbata (L.) F.H. Wigg. Dry Extracts Promote Apoptosis and DNA Damage in Human Blood Cells through Enhancing ROS Levels. Antioxidants, 10(8), 1171.

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Corneal and Conjunctival Epithelial Cell Culture

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Human corneal epithelial cells (HCE-2) [50B1] (ATCC CRL-11135) were obtained from ATCC company (American Type Culture Collection, Manassas, USA). Human conjunctival epithelial cells (HConEC) and corneal epithelium cell medium were provided by Innoprot (Derio, Bizkaia, Spain). Keratinocyte serum-free medium, bovine pituitary extract (BPE), and epidermal growth factor (EGF) were purchased from Gibco-BRL (San Giuliano Milanese, MI, Italy). Hydrocortisone, insulin, fibronectin, bovine collagen type I, bovine serum albumin (BSA), penicillin/streptomycin (P/S), bovine fetal serum, poly-l-lysine, benzalkonium chloride (BAK), and 3-(4,5-dimethylthiazol-2-yl)-25-diphenyltetrazolium bromide (MTT) were provided by Sigma (St Louis, MO, USA). The cytotoxicity detection kit (lactate dehydrogenase, LDH) was purchased from Roche Diagnostics (Basel, Switzerland).
Oftasecur and all its constituents were a kind gift from OFFHEALTH S.p.a. (Florence, Italy).
Institutional review board approval was not required at our institute for the use of immortalized cell lines.

Mencucci R., Ghelardi E., Celandroni F., Mazzantini C., Vecchione A., Pellegrini-Giampietro D.E., Favuzza E, & Landucci E. (2022). Antiseptics and the Ocular Surface: In Vitro Antimicrobial Activity and Effects on Conjunctival and Corneal Epithelial Cells of a New Liposomal Ocular Spray Containing Biosecur® Citrus Extract. Ophthalmology and Therapy, 11(3), 1067-1077.

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