Cell culture reagents

Manufactured by Thermo Fisher Scientific

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Cell culture reagents are a set of specialized solutions and compounds used to support the growth, maintenance, and manipulation of cells in laboratory settings. These reagents provide the necessary nutrients, growth factors, and environmental conditions required for cell cultures to thrive.

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Cell culture media and reagents (47 citations) Reagents for cell culture (18 citations) Culture reagents (12 citations) Cell culture-related reagents (12 citations) Total exosome isolation reagent from cell culture media (9 citations) Cell culture medium and reagents (6 citations) Other cell culture reagents (4 citations) Cell culture and transfection reagents (4 citations) Total Exosome Isolation Reagent for Cell Culture Media kit (4 citations) Primary cell culture reagents and plates (2 citations)

689 protocols using cell culture reagents

Cell Culture Protocols for HEK293T, HEK293, and CHO-K1 Cells

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HEK293T cells (CRL-3216, ATCC, Manassas, VA, USA) and HEK293 cells (catalog no. 103, NIH AIDS Reagent Program, Germantown, MD, USA) were grown in Dulbecco’s modified Eagle’s medium (DMEM) containing high glucose (4.5 g/L), 2 mM l-glutamine, 10% heat-inactivated fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin. Cell culture reagents were purchased from Gibco (Invitrogen, Carlsbad, CA, USA). CHO-K1 cells (CCL-61, ATCC, Manassas, VA, USA) were grown in Roswell Park Memorial Institute (RPMI) 1640 medium containing 2 mM l-glutamine, 10% heat-inactivated FBS, 100 U/mL penicillin, and 100 μg/mL streptomycin. Cell culture reagents were purchased from Gibco (Invitrogen, Carlsbad, CA, USA)

Bae D.H., Marino M., Iaffaldano B., Fenstermaker S., Afione S., Argaw T., McCright J., Kwilas A., Chiorini J.A., Timmons A.E, & Reiser J. (2020). Design and Testing of Vector-Producing HEK293T Cells Bearing a Genomic Deletion of the SV40 T Antigen Coding Region. Molecular Therapy. Methods & Clinical Development, 18, 631-638.

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Magnetic Glycan-Based Affinity Assay

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Wheat germ agglutinin (WGA) was purchased from Vector Laboratories (Burlingame, CA, USA). Fluorescein isothiocyanate (FITC) was obtained from Sigma-Aldrich (St. Louis, MO, USA). Magnetic nanoparticles (Nano-screen-fluidMAG/B-ARA), Magnetic separator (MagnetoPURE PM-20), Magnetic 96-well format (MagnetoFACTOR-96 plate) and CombiMAG-1000 particles were from Chemicell (Berlin, Germany). Sialic acid (N-acetyl-5-neuraminic-acid, Neu5Ac), was from Santa Cruz Biotechnology (San Diego, CA, USA). FITC BrdU Apoptosis Detection Kit was from BD Biosciences (San Jose, CA, USA). FITC-fluorescein conjugated anti-digoxigenin, MES (2-[N-Morpholino] ethanesulfonic acid) and EDC (1-ethyl-3-[3-dimethylaminoproply] carbodiimide) were from Sigma-Aldrich (Carlsbad, CA, USA). Phosphate-buffered saline (PBS, 137 mM sodium chloride, 2.7 mM potassium chloride, 4.3 mM disodium phosphate, 1.4 mM monopotassium phosphate, pH 7.5) was obtained from Technova (Hollister, CA, 95023 CA USA). Cell culture reagents were from Life Technologies Corporation (San Diego, CA, USA); all other chemicals were of analytical grade and purchased from Sigma-Aldrich.

AlSadek D.M., Badr H.A., Al-Shafie T.A., El-Bahr S.M., El-Houseini M.E., Djansugurova L.B., Li C.Z, & Ahmed H. (2017). Cancer cell death induced by nanomagnetolectin. European journal of cell biology, 96(6), 600-611.

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Rat Embryonic Fibroblast Cultivation

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All cell culture reagents were obtained from Life Technologies, Darmstadt, Germany, unless stated otherwise. Rat embryonic fibroblasts (REF52) were cultured in DMEM containing 1% penicillin/streptomycin and 10% fetal bovine serum (FBS). Cells were grown in T25 tissue culture flasks in a humidified atmosphere containing 5% CO₂ and passaged either every 3–4 d or before reaching confluency. For passaging, cells were rinsed with phosphate-buffered saline (PBS) and incubated with 1 ml trypsin/EDTA at 37°C for 5 min until detachment. Trypsin was inactivated by adding 9 ml of growth medium, and cells were diluted according to their growth rate.

Gudzenko T, & Franz C.M. (2015). Studying early stages of fibronectin fibrillogenesis in living cells by atomic force microscopy. Molecular Biology of the Cell, 26(18), 3190-3204.

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Characterization of JDP2 Protein Interactions

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All cell culture reagents were purchased from Life Technologies (Carlsbad, CA, USA). Antibodies against JDP2, MC2R, and β-Actin (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), SUMO1 and SUMO3 (Active motif, Carsbad, CA, USA), NR5A1 (Upstate Biochemistry Inc., Charlottsville, VA, USA), and FLAG (Sigma, St. Louis, MO, USA) were purchased commercially. Luciferase activity was measured using the Dual Luciferase Assay System (Promega, Madison, WI, USA). GST-JDP2 recombinant proteins were purchased from Abnova (Taipei, Taiwan).

Wang C.M., Wang R.X., Liu R, & Yang W.H. (2017). Jun Dimerization Protein 2 Activates Mc2r Transcriptional Activity: Role of Phosphorylation and SUMOylation. International Journal of Molecular Sciences, 18(2), 304.

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Breast Cancer Cell Line Analysis

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DMSO, E2 and TAM were obtained from Sigma-Aldrich (St. Louis, MO USA). RAL (Evista^®, Eli Lilly and Company, Indianapolis, IN USA) was purchased from the University of Illinois at Chicago Hospital Pharmacy. Cell culture reagents were obtained from Life Technologies (Carlsbad, CA USA). Tissue culture plasticware was purchased from Becton-Dickinson (Franklin Lakes, NJ USA). The following antibodies were used: rabbit polyclonal PKCα (C-20, Santa Cruz Biotechnology, Dallas, TX USA), rabbit polyclonal ERα (SP1, Lab Vision, Thermo Scientific, Kalamazoo, MI USA), mouse monoclonal β-actin (Sigma, St. Louis, MO USA), anti-rabbit Alexa Fluor 488 (Life Technologies, Carlsbad, CA USA) and anti-mouse Cy3 (Jackson Immunoresearch Laboratories, West Grove, PA).

Molloy M.E., Perez White B.E., Gherezghiher T., Michalsen B.T., Xiong R., Patel H., Zhao H., Maximov P.Y., Jordan V.C., Thatcher G.R, & Tonetti D.A. (2014). Novel Selective Estrogen Mimics for the Treatment of Tamoxifen-Resistant Breast Cancer. Molecular cancer therapeutics, 13(11), 2515-2526.

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PCTAIRE-tide Peptide Substrate for CDK16 Kinase Assays

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PCTAIRE-tide (PKSPKARKKL) peptide substrate for CDK16 kinase assays was synthesised by GL Biochem. [^γ-32P]ATP was obtained from PerkinElmer. Cell culture reagents were obtained from Life Technologies. Preclinical kinase inhibitors were obtained from Calbiochem. Clinical kinase inhibitors were obtained from the FIMM drug collection. Published kinase inhibitor set (PKIS) compounds were a gift from William Zuercher (GlaxoSmithKline). P81 paper was obtained from Whatman. Unless otherwise indicated, all other reagents were obtained from Sigma.

Dixon-Clarke S.E., Shehata S.N., Krojer T., Sharpe T.D., vonDelft F., Sakamoto K, & Bullock A.N. (2017). Structure and inhibitor specificity of the PCTAIRE-family kinase CDK16. Biochemical Journal, 474(5), 699-713.

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Photofrin-Doxycycline Combination Therapy

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Photofrin® (75 mg/vial) was obtained from Union Med. Group Limited Company, Hong Kong, China. Doxycycline (Dox) was supplied by Abcam. Puromycin was provided by Abcam. Cell culture reagents were obtained from Life Technology. Other reagents were all supplied by Sigma-Aldrich unless otherwise stated.

Wu D.P., Bai L.R., Lv Y.F., Zhou Y., Ding C.H., Yang S.M., Zhang F., Wang Y.Y., Huang J.L, & Yin X.X. (2019). A Novel Role of Connexin 40-Formed Channels in the Enhanced Efficacy of Photodynamic Therapy. Frontiers in Oncology, 9, 595.

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Comprehensive Metabolic Assay Protocol

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The HFD formula fodder (D12492) was obtained from Guangdong Medical Laboratory Animal Center, (Guangzhou, China). All cell culture reagents were obtained from GibcoBRL (Life Technologies, Inc., Grand Island, NY). mRFP-GFP-LC3 adenoviruses were purchased from Hanbio Co. (Shanghai, China). TRIzol reagent and SuperScript III First-Strand Synthesis Supermix were from Invitrogen (Carlsbad, California). SYBR Green PCR Master Mix was from Applied Biosystems (Foster City, California). TG reagents were from Nanjing Jiancheng Biocompany (Nanjing, China). sEH rabbit mAb was kindly donated by Dr. Bruce D. Hammock from University of California. Species- specific secondary antibodies were from Sigma-Aldrich. Enhanced chemiluminescent substrate was from Pierce (Rockford, IL). Polyvinylidene difluoride (PVDF) membranes were from Merck Millipore (Billerica, MA). All other chemicals and reagents were purchased from Sigma-Aldrich unless otherwise specified.

Wang L., Zhao D., Tang L., Li H., Liu Z., Gao J., Edin M.L., Zhang H., Zhang K., Chen J., Zhu X., Wang D., Zeldin D.C., Hammock B.D., Wang J, & Huang H. (2020). Soluble Epoxide Hydrolase Deficiency Attenuates Lipotoxic Cardiomyopathy via Upregulation of AMPK-mTORC Mediated Autophagy. Journal of molecular and cellular cardiology, 154, 80-91.

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Apoptosis Assay in Cell Cultures

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Dulbecco’s Modified Eagle’s Medium (DMEM) and all cell culture reagents were purchased from Life Technologies (CA, USA). Fetal bovine serum (FBS) and MTT were obtained from Himedia. FITC AnnexinV Apoptosis Detection Kit was bought from Becton Dickinson (NJ, USA). All other chemicals were of analytical grade. A colorimetric assay kit was used to measure caspase-3 activity (R&D Systems, Inc., Minneapolis, MN, USA). Enzyme-linked immunonosorbent assay (ELISA) kits were used to measure Bax and Bcl-2 protein levels (Elabscience, Houston, TX, USA). Ethanol solution (60%), for extraction procedures, was prepared using ethanol PA from Synth (Brazil). Sodium carbonate and aluminum chloride were obtained from Dinâmica (Brazil) and Synth, respectively, while Folin-Ciocalteu was obtained from Merck^®. Gallic acid and quercetin standards were purchased from Sigma (Annapolis, MD, USA). Methanol was used for UPLC/MS procedures, acetonitrile and formic acid were HPLC grade from Merck^®, and sodium formate was prepared from sodium hydroxide (Dinâmica, Brazil) and formic acid (Merck^®).

Marcucci M.C., Oliveira C.R., Spindola D., Antunes A.A., Santana L.Y., Cavalaro V., Costa I.B., de Carvalho A.C., Veiga T.A., Medeiros L.S., dos Santos Zamarioli L., Gonçalves C.P., Santos M.F., Grecco S.S., Suzuki V.Y., Ferreira L.M, & Garcia D.M. (2022). Molecular Dereplication and In Vitro and In Silico Pharmacological Evaluation of Coriandrum sativum against Neuroblastoma Cells. Molecules, 27(17), 5389.

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Reagent Optimization for Cell Culture

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All the reagents, unless otherwise stated, were from Merck KGaA (Darmstadt, Germany). Cell culture reagents were from Life Technologies (GibcoBRL, Gaithersburg, MD, USA).

Costanzi E., Romani R., Scarpelli P, & Bellezza I. (2020). Extracellular Vesicles-Mediated Transfer of miRNA Let-7b from PC3 Cells to Macrophages. Genes, 11(12), 1495.

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