F 12k media

Manufactured by American Type Culture Collection

Sourced in United States

F-12K media is a cell culture medium designed for the growth and maintenance of a variety of cell lines. It provides essential nutrients and growth factors required for cell proliferation and survival in controlled in vitro environments.

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Lab products found in correlation

16 protocols using f 12k media

Cell Culture of M17 and SH-SY5Y

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SK-N-BE(2)-M17 (M17) cells (ATCC, Manassas, VA) and SH-SY5Y cells (ATCC) were grown in Eagles minimal essential media (MEM) and F12K media (ATCC) at 1:1 ratio supplemented with 10% fetal bovine serum (SAFC Biosciences, Lenexa, KS) and 1% penicillin/streptomycin (Life Technologies) at 37°C with 5% CO2 in a humidified incubator.

Slone S.R., Lavalley N., McFerrin M., Wang B, & Yacoubian T.A. (2015). Increased 14-3-3 phosphorylation observed in Parkinson’s disease reduces neuroprotective potential of 14-3-3 proteins. Neurobiology of disease, 79, 1-13.

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Respiratory Syncytial Virus Infection Assay

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A549 cells were grown in F-12K media (ATCC) containing 10% FBS and were seeded in 6-well plates and infected with rRSV or rRSVflag(2)L in triplicate at an MOI of 0.01. After 1 hr absorption, media was replaced with 1 mL of serum containing media. At indicated timepoints, media overlay was snap-frozen and stored at -80°C and subsequently replaced every 24 hrs for 7 days. Virus titer was determined by plaque assay.

Blanchard E.L., Braun M.R., Lifland A.W., Ludeke B., Noton S.L., Vanover D., Zurla C., Fearns R, & Santangelo P.J. (2020). Polymerase-tagged respiratory syncytial virus reveals a dynamic rearrangement of the ribonucleocapsid complex during infection. PLoS Pathogens, 16(10), e1008987.

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Cell Line Culturing Protocol

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CT26WT (CRL-2638; American Type Culture Collection [ATCC]), CT26LacZ (CRL-2639; ATCC), TC-1 (provided by T.C. Wu of Johns Hopkins University), and 4T1 (CRL-2538; ATCC) were maintained in RPMI 1640 medium (Thermo Scientific, MA, USA) supplemented with 10% fetal bovine sera (FBSs). L929 (CCL-1; ATCC), B16-F1 (CRL-6323; ATCC), Vero (CCL-81; ATCC), HeLa (CCL-2; ATCC), U-87 MG (HTB-14; ATCC), PANC-1 (CRL-146; ATCC), and A549 (CCL-185; ATCC) were maintained in DMEM (Thermo Scientific, MA, USA) supplemented with 10% FBS. SKOV3 (HTB-77; ATCC) cell lines were grown in McCoy’s 5a medium (30-2007; ATCC) supplemented with 10% FBS. PC3 (CCL-2; ATCC) cell lines were maintained in F12K media (30-2004; ATCC) supplemented with 10% FBS. EMT-6 (CRL-2755; ATCC) cell lines were maintained in Weymouth’s media (MD-7193; ATCC) supplemented with 15% FBS. Cell lines were purchased from and verified by the ATCC (Manassas, VA, USA). All media were supplemented with 100 U/mL penicillin and 100 μg/mL streptomycin, and cells were incubated in a humidified 37°C incubator with 5% CO₂.

Maroun J.W., Penza V., Weiskittel T.M., Schulze A.J, & Russell S.J. (2020). Collateral Lethal Effects of Complementary Oncolytic Viruses. Molecular Therapy Oncolytics, 18, 236-246.

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A549 Lung Cancer Cell Culture

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A549 human
lung adenocarcinoma
cells (ATCC CCL-185) were cultured in F-12K media (ATCC 30-2004) supplemented
with 10% fetal bovine serum (Atlanta Biologicals) and 1% penicillin/streptomycin
(Sigma-Aldrich) and maintained at 37 °C and 5% CO₂ in a humidified incubator. Cell authentication experiments using
fluorescent antibodies confirmed that the cells expressed the α_vβ₅ integrin receptor.^{31 (link)}

Gamage R.S., Li D.H., Schreiber C.L, & Smith B.D. (2021). Comparison of cRGDfK Peptide Probes with Appended Shielded Heptamethine Cyanine Dye (s775z) for Near Infrared Fluorescence Imaging of Cancer. ACS Omega, 6(44), 30130-30139.

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Comparative Analysis of Prostate and Breast Cancer Cell Lines

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Four prostate cancer cell lines (VCaP, LNCaP, PC-3 and PC-3M) and two breast cancer cell lines (SKBR3 and MDAMB-231) were used in our analysis. VCaP, SKBR3, and MDAMB231 were obtained from American Type Culture Collection (ATCC). VCaP cells were cultured in Dulbecco’s Modified Eagles’s Medium (DMEM, ATCC). SKBR3 cells were cultured in McCoy’s 5a Medium Modified (ATCC) and MDA-MB-231 cells were cultured in Leibovitz’s L-15 medium (ATCC). LNCaP, PC-3 and PC-3M cell lines were obtained from Dr. Alison Allan’s research group at London Health Sciences, London, ON, Canada. LNCaP and PC-3M cells were cultured in RPMI-1640. PC-3 cells were maintained in F12K media (ATCC). All media were supplemented with 10% fetal bovine serum (FBS). Cells were cultured at 37 °C and 5% CO₂.

Kermanshah L., Poudineh M., Ahmed S., Nguyen L.N., Srikant S., Makonnen R., Cantu F.P., Corrigan M, & Kelley S.O. (2018). Dynamic CTC Phenotypes in Prostate Cancer Metastatic Models Visualized Using Magnetic Ranking Cytometry. Lab on a chip, 18(14), 2055-2064.

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Culturing Colorectal Cancer Cell Lines

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The human colorectal cancer cell lines HT-29 (grade 1) and LoVo (grade 4) as well as CCD-18Co cells representing the normal colonic epithelium were obtained from ATCC (Manassas, VA, USA). HT-29 cells were cultured with McCoy’s 5A (ATCC), LoVo cells with F12-K media (ATCC), CCD18-Co with Eagle’s minimum essential medium (Lonza, Basel, Switzerland) with 1% sodium pyruvate and 1% nonessential amino acids (all from Sigma-Aldrich, St. Louis, MO, USA). All media contained 1% l-glutamine and penicillin-streptomycin solution and 10% fetal bovine serum (Sigma-Aldrich). Cell cultures were provided in 5% CO 2 at 37 °C and 95% humidity. The medium was changed twice a week, and cells were passaged at approximately 70% confluence and trypsinized with 0.25% trypsin–EDTA solution (Sigma-Aldrich).

Buzalewicz I., Mrozowska M., Kmiecik A., Kulus M., Haczkiewicz-Leśniak K., Dzięgiel P., Podhorska-Okołów M, & Zadka Ł. (2022). Quantitative Phase Imaging Detecting the Hypoxia-Induced Patterns in Healthy and Neoplastic Human Colonic Epithelial Cells. Cells, 11(22), 3599.

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Viability and Apoptosis Assays for Common Cancer Cell Lines

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All cell lines were obtained from ATCC. A549 lung carcinoma cells were cultured in F-12K media (ATCC). K562 chronic myelogenous leukemia cells and THP-1 acute monocytic leukemia cells were cultured in RPMI media (Gibco). MDA-MB-231 breast cancer cells were cultured in Dulbecco's modified Eagle's medium (Gibco). All media were supplemented with 10% fetal bovine serum (HyClone) and penicillin/streptomycin (pen/strep; Gibco). RealTime-Glo MT Cell Viability Assay, CellTiter-Glo, CellTiter-Fluor, and Caspase-Glo 3/7 were obtained from Promega Corporation. White 384-well assay plates were purchased from Corning, Inc. All reagents were purchased through Sigma, unless otherwise noted.

Duellman S.J., Zhou W., Meisenheimer P., Vidugiris G., Cali J.J., Gautam P., Wennerberg K, & Vidugiriene J. (2015). Bioluminescent, Nonlytic, Real-Time Cell Viability Assay and Use in Inhibitor Screening. Assay and Drug Development Technologies, 13(8), 456-465.

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Establishing PC-12 Cell Culture

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We purchased F-12K media, trypsin-EDTA solution, horse serum, fetal bovine serum, and the rat pheochromocytoma (PC) PC-12 cell line from ATCC (Manassas, VA, USA). Poly-D-lysine was acquired from Sigma (St. Louis, MO, USA). An Adenosine 5′-triphosphate (ATP) Bioluminescent Assay Kit was bought from Promega (Madison, WI, USA) (Supplementary Table S1).

Modi J.P., Shen W., Menzie-Suderam J., Xu H., Lin C.H., Tao R., Prentice H.M., Schloss J, & Wu J.Y. (2023). The Role of NMDA Receptor Partial Antagonist, Carbamathione, as a Therapeutic Agent for Transient Global Ischemia. Biomedicines, 11(7), 1885.

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A549 cell IFN-gamma treatment protocol

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A549 cells (catalog #CCL-185, ATCC, Manassas VA) were grown in F-12K media (catalog #30–2004, ATCC, Manassas VA) plus 10% fetal bovine serum (catalog #35-015-CV, Corning, Amsterdam Netherlands). Cell cultures were maintained at a density below the recommendation by ATCC of 6 x 10⁴ cells/cm². At passage number three, cells were either treated with IFN γ (catalog #PHC-4031, ThermoFisher Scientific, Waltham, MA) at 100 ng/mL for 48 hours or left untreated. After 48 hours the cells were counted, harvested, and RNA extracted.

O’Rourke D., Wang D., Sanchez-Garcia J.F., Cusano M.P., Miller W., Cai T., Scheuenpflug J, & Feng Z. (2021). Fit-for-purpose quantitative liquid biopsy based droplet digital PCR assay development for detection of programmed cell death ligand-1 (PD-L1) RNA expression in PAXgene blood samples. PLoS ONE, 16(5), e0250849.

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Differentiation of SH-SY5Y Neuroblastoma Cells

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Naïve SH-SY5Y human neuroblastoma cells (ATCC, CRL-2266) were maintained in T-75 flasks (Corning, 430641) at 37°C and 5% CO₂ in Minimum Essential Media (Cellgro, 10-010-CV) and F12-K media (ATCC, 30–2004) supplemented with 0.5% sodium pyruvate (Cellgro, 25-000-CI), 0.5% non-essential amino acids (Cellgro, 25-025-CI), 1% penicillin/streptomycin (Invitrogen, 15140-122), and 10% heat-inactivated fetal bovine serum (FBS; Thermo Scientific, SH30109.03). Naïve cells received full media change every two days. Prior to their use in experiments, naïve SH-SY5Y cells were differentiated for seven days to a post-mitotic state in 10% FBS media supplemented with 10 μM retinoic acid (RA; Sigma, R2625). Complete RA-supplemented media was replenished every two days.

Mangieri L.R., Mader B.J., Thomas C.E., Taylor C.A., Luker A.M., Tse T.E., Huisingh C, & Shacka J.J. (2014). ATP6V0C Knockdown in Neuroblastoma Cells Alters Autophagy-Lysosome Pathway Function and Metabolism of Proteins that Accumulate in Neurodegenerative Disease. PLoS ONE, 9(4), e93257.

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