Fetal calf sera

Manufactured by Thermo Fisher Scientific

Sourced in United States

Fetal calf sera is a complex mixture of proteins, growth factors, and other nutrients derived from the blood of bovine fetuses. It is commonly used as a supplement in cell culture media to provide essential components for the growth and maintenance of various cell lines.

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8 protocols using fetal calf sera

Extracellular Matrix Signaling Assay

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Dulbecco’s Modified Eagle’s Media (DMEM), fetal calf sera, trypsin solution, and penicillin/streptomycin were purchased from Invitrogen (Carlsbad, CA). [γ-³²P]ATP was obtained from New England Nuclear Life Science Products (Boston, MA). CCN1 antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Type I collagen antibody was purchased from SouthernBiotech (Birmingham, AL). Total and phosphorylated c-Jun antibodies were purchased from Transduction Laboratories (San Diego, CA) and Santa Cruz Biotechnology (Santa Cruz, CA), respectively. β-actin antibody was purchased from Sigma Chemical (St. Louis, MO). All other reagents were purchased from Sigma Chemical.

Qin Z., Robichaud P., He T., Fisher G.J., Voorhees J.J, & Quan T. (2014). Oxidant Exposure Induces Cysteine-Rich Protein 61 (CCN1) via c-Jun/AP-1 to Reduce Collagen Expression in Human Dermal Fibroblasts. PLoS ONE, 9(12), e115402.

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EGCG Treatment and Boceprevir Analysis

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Dulbecco’s modified Eagle’s medium (DMEM), glutamax-I, goat, and fetal calf sera were purchased from Invitrogen (Carlsbad, CA, USA). 4′,6-Diamidino-2-phenylindole (DAPI) was from Molecular Probes (Life technologies). EGCG was from Calbiochem (Merck Chemicals, Darmstadt, Germany). Stocks were resuspended in dimethyl sulfoxide (DMSO) at 0.5 M for EGCG and 250 mg/mL for plant extracts. Boceprevir was kindly provided by Philippe Halfon (Hôpital Européen, Laboratoire Alphabio, Marseille, France).

Galani B.R., Sahuc M.E., Njayou F.N., Deloison G., Mkounga P., Feudjou W.F., Brodin P., Rouillé Y., Nkengfack A.E., Moundipa P.F, & Séron K. (2015). Plant extracts from Cameroonian medicinal plants strongly inhibit hepatitis C virus infection in vitro. Frontiers in Microbiology, 6, 488.

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Activation and HIV Infection of PBMC

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Peripheral blood mononuclear cells were isolated from whole blood samples by using a Lymphoprep™ (Stemcell) density gradient and were frozen until use. To activate PBMC prior to infection with HIV, cells were thawed and cultured for 3 days in phytohemagglutinin (PHA, 5 µg/ml) containing R20/50 medium. R20/50 medium consisted of Roswell Park Memorial Institute (RPMI) 1640 (Gibco) supplemented with 2 mM l-glutamine (Gibco), 100 U/ml penicillin, 100 µg/ml streptomycin (Gibco), 20% of fetal calf sera (Invitrogen), and 50 U/ml of interleukin 2 (IL-2, Roche). The 3-days PHA-blasts were used as target cells in infection assays either unmodified or pre-incubated overnight with BAGN. PBMC were obtained from HIV sero-negative and from HIV-infected, treatment naïve, donors. Before obtaining any samples, donors signed informed consent forms approved by the Ethics Committee of the Hospital Universitari Germans Trias i Pujol (Badalona, Spain).

Olvera A., Martinez J.P., Casadellà M., Llano A., Rosás M., Mothe B., Ruiz-Riol M., Arsequell G., Valencia G., Noguera-Julian M., Paredes R., Meyerhans A, & Brander C. (2018). Benzyl-2-Acetamido-2-Deoxy-α-d-Galactopyranoside Increases Human Immunodeficiency Virus Replication and Viral Outgrowth Efficacy In Vitro. Frontiers in Immunology, 8, 2010.

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Collagen-based Cell Culture Protocol

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Dulbecco's Modified Eagle's Media (DMEM), fetal calf sera, trypsin solution, and penicillin/streptomycin were obtained from Invitrogen Life Technology (Carlsbad, CA, USA). [γ‐³²P]ATP was obtained from New England Nuclear Life Science Products (Boston, MA, USA). Rat tail type I collagen was purchased from BD Biosciences (Palo Alto, CA, USA). TGF‐β1 was purchased from R&D Systems (Minneapolis, MN, USA). Latrunculin‐A was purchased from Enzo Life Sciences (Farmingdale, NY, USA). All other reagents were purchased from Sigma Chemical Company (St. Louis, MO, USA).

Fisher G.J., Shao Y., He T., Qin Z., Perry D., Voorhees J.J, & Quan T. (2015). Reduction of fibroblast size/mechanical force down‐regulates TGF‐β type II receptor: implications for human skin aging. Aging Cell, 15(1), 67-76.

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Cultivation of Murine Lung Epithelial Cells

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We purchased the murine lung epithelial cell line (MLE-12) from the Institute of Stem Cell Research within the Chinese Academy of Sciences (Shanghai, China). MLE-12 cells were cultured in DMEM-f12 (Gibco, Grand Island, NY, USA) supplemented with 10% heat-inactivated fetal calf sera (Gibco), 1% penicillin, and streptomycin (Millipore, Waltham, MA, USA) in a 37°C incubator with 5% CO₂.

Wang Y.M., Gong F.C., Qi X., Zheng Y.J., Zheng X.T., Chen Y., Yang Z.T., Q.i.n.g.-.Y.e., Mao E.Q, & Chen E.Z. (2022). Mucin 1 Inhibits Ferroptosis and Sensitizes Vitamin E to Alleviate Sepsis-Induced Acute Lung Injury through GSK3β/Keap1-Nrf2-GPX4 Pathway. Oxidative Medicine and Cellular Longevity, 2022, 2405943.

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Macrophage Polarization and Netrin-1 Expression

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Human monocytic cell line THP-1 cells and rat alveolar macrophage-derived cell line NR8383 cells were purchased from the Stem Cell Bank at Chinese Academy of Sciences. The cells were cultured with Dulbecco’s Modified Eagle Medium/F-12 containing 15% fetal calf sera (Gibco). The THP-1 cells were induced to the macrophages (M0) by phorbol-12-myristate-13-acetate (PMA, 20 ng/mL, Sigma). Subsequently, the differentiated human THP-1 macrophages and NR8383 cells were treated with lipopolysaccharide (LPS, 20 ng/mL, Sigma) and interferon gamma (IFN-γ, 20 ng/mL, PeproTech) to further induce M1 phenotype macrophages. Next, expression levels of the M1 phenotype markers tumor necrosis factor alpha (TNF-α), interleukin (IL)-1β, IL-6, MCP-1 and nitric oxide synthase 2 (iNOS) were assessed at 1, 3, 6, 12, 24 and 48 hours after stimulation. Meanwhile, mRNA and protein expression levels of netrin-1 as well as secreted netrin-1 levels in cell supernatants were measured using qRT-PCR, Western blot and ELISA respectively.

Ding S., Guo X., Zhu L., Wang J., Li T., Yu Q, & Zhang X. (2021). Macrophage-derived netrin-1 contributes to endometriosis-associated pain. Annals of Translational Medicine, 9(1), 29.

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Cell Viability Assay Reagents

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Dimethylsulfoxide (DMSO), L-glutamine, adenosine, gentamicin, adenosine, HEPES, hemin and Amphotericin B, were bought from Gibco (Gibco, Life Technologies GmbH, Karlsruhe, Germany) and Sigma (Darmstadt, Germany). 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was obtained from Sigma-Aldrich (Deisenhofen, Germany). Schneider’s Drosophila medium, M199 medium, Dulbecco’s modified Eagle’s medium (DMEM) phenol red-free, RPMI-1640, Fetal Calf Sera (FCS) were supplied from Gibco (Gibco, Invitrogen Corporation, Carlsbad, CA).

Badirzadeh A., Heidari-Kharaji M., Fallah-Omrani V., Dabiri H., Araghi A, & Salimi Chirani A. (2020). Antileishmanial activity of Urtica dioica extract against zoonotic cutaneous leishmaniasis. PLoS Neglected Tropical Diseases, 14(1), e0007843.

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Reagents for Molecular Biology Techniques

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All solutions were prepared using MilliQ ultrapure (Milli-QSystem, Millipore, Molsheim, France) and non-pyrogenic water to avoid surface-active impurities. G418, and Sodium dodecyl sulfate (SDS) were purchased from Sigma-Aldrich (Sigma, Deisenhofen, Germany). The material for PCR, enzymatic digestion and agarose gel electrophoresis were acquired from Roche Applied Sciences (Mannheim, Germany). Cell culture reagents including M199 medium, HEPES, L-glutamine, adenosine, hemin, gentamicin, DMEM and Schneider were purchased from Sigma (Darmstadt, Germany) and Gibco (Gibco, Life Technologies GmbH, Karlsruhe, Germany), respectively. Fetal Calf Sera (FCS) was purchased from Gibco (Gibco, Life Technologies GmbH, Karlsruhe, Germany). All cytokine kits were purchased from DuoSet R & D kits, (Minneapolis, USA).

Zahedifard F., Gholami E., Taheri T., Taslimi Y., Doustdari F., Seyed N., Torkashvand F., Meneses C., Papadopoulou B., Kamhawi S., Valenzuela J.G, & Rafati S. (2014). Enhanced Protective Efficacy of Nonpathogenic Recombinant Leishmania tarentolae Expressing Cysteine Proteinases Combined with a Sand Fly Salivary Antigen. PLoS Neglected Tropical Diseases, 8(3), e2751.

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