Dmed f12

Manufactured by Thermo Fisher Scientific

Sourced in United States

The DMED/F12 is a laboratory equipment product designed for cell culture applications. It serves as a culture medium to support the growth and maintenance of various cell types. The product combines the key components necessary for cell proliferation and viability.

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DMED-F12 medium (5 citations)

8 protocols using dmed f12

Glioblastoma Cell Culture and Transduction

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U-251MG and U-87MG cell lines were purchased from China Infrastructure of Cell Line Resources (Kunming, China). They were cultured in DMEM supplemented with 10% FBS, 100 U/mL penicillin and 100 mg/mL streptomycin (Life Technologies, USA). The GSC-3# and GSC12# cell lines, isolated from primary glioblastoma tissues, were previously described [29 (link)]. U-251MG cells and U-87MG cells were transduced with lentivirus vectors expressing luciferase, which were named U251-luc and U87-luc respectively. U-251MG-derived CSC spheres were cultured from U251-luc cells in the medium containing serum-free DMED/F12 supplemented with 20 ng/mL EGF, 20 ng/mL bFGF and 1x B27 (all from Life Technologies, USA). All cells were grown in a humidified incubator with 5% CO₂ at 37 ℃.

Wang S., Wei W., Yuan Y., Sun B., Yang D., Liu N, & Zhao X. (2023). Chimeric antigen receptor T cells targeting cell surface GRP78 efficiently kill glioblastoma and cancer stem cells. Journal of Translational Medicine, 21, 493.

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Cancer Stem Cell Culture Protocols

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Two non-small-cell lung cancer (NSCLC) cell lines, A549 and H1299, and 293T cells were purchased from the Chinese National Infrastructure of Cell Line Resources (Kunming, China). Cells were cultured in a complete medium composed of DMEM with 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin (Life Technologies, Carlsbad, CA, USA). A549- or H1299-derived cancer stemlike cells (CSCs), named A549 CSCs or H1299 CSCs, were cultured with a conditioned stem-cell medium of serum-free DMED/F12, supplemented with 20 ng/mL EGF, 20 ng/mL bFGF and 1× B27 (all supplements purchased from Life Technologies, Carlsbad, CA, USA). All cells were grown in a humidified incubator with 5% CO₂ at 37 °C.

Wang S., Wei W., Yuan Y., Guo J., Liang D, & Zhao X. (2024). Cell-Surface GRP78-Targeted Chimeric Antigen Receptor T Cells Eliminate Lung Cancer Tumor Xenografts. International Journal of Molecular Sciences, 25(1), 564.

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Isolation and Expansion of Rat BMSCs

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BMSCs were isolated from the femurs and tibias of 5 weeks old SD rats [42 (link)]. Concisely, euthanized SD rats were sterilized by dipping in 75% ethanol for 15 min, then femurs and tibias of SD rats were separated in a sterile environment and bone marrow were flushed out by using DMED/F12 (Gibco, New York, USA) added 10% Fetal Bovine Serum (FBS) (Gibco) and 1% penicillin/streptomycin (Boster). The culture medium was replaced every 2 days. After reaching 90% clustering, BMSCs were subcultured by digesting with 0.25% trypsin. Third passage of BMSCs was chosen for the following experiments.

Guo J., Ren R., Yao X., Ye Y., Sun K., Lin J., Wang G., Guo F., Xiao J, & Xu T. (2020). PKM2 suppresses osteogenesis and facilitates adipogenesis by regulating β-catenin signaling and mitochondrial fusion and fission. Aging (Albany NY), 12(4), 3976-3992.

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Cell Line Cultivation and MTT Assay

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Human colorectal adenocarcinoma cell line HT-29, human hepatocarcinoma cell line HepG2, and mouse embryonic fibroblasts (NIH3T3) were provided from the National Cell Bank of Pasture Institute of Iran (NCBI) and Iranian Biological Resource Center (IBRC). Dulbecco's modified eagle’s medium (DMEM), DMED/F12, FBS (Fetal Bovine Serum), trypsin–EDTA, and penicillin G/streptomycin were purchased from Gibco TM (Gibco-BRL, Rockville, IN, USA). The 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium (MTT) was obtained from Sigma‑Aldrich (Saint Louis, Missouri, USA). Other chemicals were supplied by Merck (Darmstadt, Germany) and Sigma-Aldrich (St Louis, MO, USA).

Koopaei N.N., Shademani M., Yazdi N.S., Tahmasvand R., Dehbid M., Koopaei M.N., Azizian H., Mousavi Z., Almasirad A, & Salimi M. (2022). Design and synthesis of novel ureido and thioureido conjugated hydrazone derivatives with potent anticancer activity. BMC Chemistry, 16(1), 81.

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Cell Culture Conditions for Transfection

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HEK293T and HeLa cells were cultured in DMEM (Sigma) supplemented with 10% FBS (Sigma), 1% GlutaMAX (Thermo Fisher Scientific), and 1% gentamicin (Thermo Fisher Scientific). The cells were transfected with expression plasmids with the use of jetPEI (Polyplus Transfection). IMCD cells were cultured in DMED/F12 (1:1) (Gibco 11320-033) with 10% FBS. For RBNS analysis, the human embryonic kidney cell line 293FT (Invitrogen, #R70007) was cultured under a humidified atmosphere of 5% CO₂ at 37 °C in Dulbecco’s modified Eagle’s medium (DMEM) (Nacalai tesque, #08459-64) supplemented with 10% fetal bovine serum (FBS) (Biosera, #554-02155, lot #10259), 2 mM L-glutamine (Thermo Fisher Scientific, #25030-081), 1 × MEM Nonessential Amino Acids (Thermo Fisher Scientific, #11140-050), and 1 mM sodium pyruvate (Sigma, #S8636-100ML).

Minegishi K., Rothé B., Komatsu K.R., Ono H., Ikawa Y., Nishimura H., Katoh T.A., Kajikawa E., Sai X., Miyashita E., Takaoka K., Bando K., Kiyonari H., Yamamoto T., Saito H., Constam D.B, & Hamada H. (2021). Fluid flow-induced left-right asymmetric decay of Dand5 mRNA in the mouse embryo requires a Bicc1-Ccr4 RNA degradation complex. Nature Communications, 12, 4071.

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Ameliorating IL-1β-induced Chondrocyte Damage

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Human MSCs (hMSCs) and human chondrocyte C28/I2 cells were acquired from ATCC (Manassas, United States) and BeNa Culture Collection (Beijing, China), respectively. Cells were cultured in DMED/F12 (Gibco, United States), containing 10% FBS (Gibco, United States) in a humidified incubator at 37°C and 5% CO₂. Cells were passaged every 2–3 days.
To establish an OA cell model, C28/I2 cells at 60–70% confluency were treated with or without 10 ng/ml human IL-1β recombinant protein (Sigma, United States) for 24 h. Following establishment of the vitro OA model, chondrocytes were treated with normal medium, hMSCs-EVs (10 μg/ml), and hMSCs^malat−1-EVs (EVs derived from lncRNA malat-1-overexpressing-hMSCs) for 24 h to investigate the effect of EVs, derived from lncRNA malat-1-overexpressing-hMSCs, on IL-1β-induced chondrocyte damage.

Pan C., Huang W., Chen Q., Xu J., Yao G., Li B., Wu T., Yin C, & Cheng X. (2021). LncRNA Malat-1 From MSCs-Derived Extracellular Vesicles Suppresses Inflammation and Cartilage Degradation in Osteoarthritis. Frontiers in Bioengineering and Biotechnology, 9, 772002.

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Culturing Human Limbal Epithelial Cells

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Human limbal rings from cadaveric corneas were obtained under the approval of the local ethical committee and declaration of Helsinki from at least 3 corneas from 3 different donors. The epithelium was separated from the underlying stroma following incubation with dispase II (Gibco, Life Technologies, USA). Cells were cultured at 37°C, 5% CO2, and 20% O2. For clonogenicity assay cells were grown in co-culture with mitomycinized growth-arrested J2-NIH3T3 cells in Green medium (60% DMEM (Gibco), 30% DMED F12 (Gibco), 10% FCII serum (Hyclone), 1mM L-Glutamine (Biological Industries), 1mM Sodium Pyruvate (Biological Industries), 0.2mM Adenine (Sigma), 5μg/ml Insulin (Sigma), 0.5μg/ml Hydrocortisone (Sigma), 10mM Choleratoxin (Sigma), 10ng/ml EGF (Peprotec)) (Rheinwald and Green, 1977 (link)) and split in 80% confluence. For efficient transfection and for controlled calcium-induced differentiation, cells were switched to defined EPIGRO medium with supplements (SCMK001, Millipore, USA) containing 1% penicillin/streptomycin and low calcium (150μM).

Altshuler A., Amitai-Lange A., Tarazi N., Dey S., Strinkovsky L., Hadad-Porat S., Bhattacharya S., Nasser W., Imeri J., Ben-David G., Abboud-Jarrous G., Tiosano B., Berkowitz E., Karin N., Savir Y, & Shalom-Feuerstein R. (2021). Discrete limbal epithelial stem cell populations mediate corneal homeostasis and wound healing. Cell stem cell, 28(7), 1248-1261.e8.

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Breast Cancer Cell Line Culture

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Breast cancer cell lines MCF-7 and T47D gained from the American Type Culture Collection (ATCC) were cultured by high-glucose (4.5 mg/ml) DMEM (HyClone, USA) with 10% serum (Tianjin Hao Yang Biological Manufacture CL, China). MCF-7 mammosphere (MCF-7 MS) and T47D mammosphere (T47D MS) were inducted and cultured by DMED-F12 (Gibco) with EGF 20 µg/L (Promega), b-FGF 10 µg/L (Promega) and 2% B27 (Gibco). All of the cells used in our research were cultured in the condition containing 5% CO 2 and 95% air at 37°C.

Li S., Ma J., Zheng A., Song X., Chen S., & Jin F. (2021). DEAD-Box Helicase 27 Enhances Stem Cell-Like Properties With Poor Prognosis in Breast Cancer.

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