Dmem f12 gibco

Manufactured by Thermo Fisher Scientific

Sourced in United States, China

DMEM-F12 GIBCO is a basal medium formulated for the culture of mammalian cells, including those derived from human, mouse, and other species. It is a combination of Dulbecco's Modified Eagle's Medium (DMEM) and Ham's F-12 Nutrient Mixture, designed to support the growth and maintenance of a variety of cell lines.

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9 protocols using dmem f12 gibco

Chick Embryo Primitive Streak Explant Culture

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Explant culture: HH3 chick embryo primitive streak is divided into six equal size portions, and the anterior primitive streak (cardiac progenitor cells) is deemed to be the second segment of explants from the cranial side (Fig. S4A). The explants of anterior primitive streaks were cultured in vitro in culture medium (DMEM-F12 GIBCO) at 37 o C and 5% CO2 [25] for 24, 48 or 72 hours. Each treatment was performed in triplicate. Approximately, 95% of the cells in the explant culture were determined to be cardiomyocytes in the control group, which shows the presence of myosin heavy chain using MF20 immunofluorescent staining as previously reported [26] .
Primary culture: Primary cardiomyocyte cultures were established from day 14 chick embryo hearts (DMEM-F12 GIBCO).
Cell culture: H9c2 cells was purchased from Guangzhou Jennio Biotech Co.,Ltd, China and cultured in culture medium (DMEM-F12 GIBCO).

Zhang J., Wang G., Liu J., Gao L.R., Liu M., Wang C.J., Chuai M., Bao Y., Li G., Li R.M., Zhang Y, & Yang X. (2018). Gut microbiota-derived endotoxin enhanced the incidence of cardia bifida during cardiogenesis. Journal of cellular physiology, 233(12).

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Choriocarcinoma Trophoblast Cell Lines Protocol

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The choriocarcinoma trophoblast cell lines, BeWo, Jar, and Jeg‐3, were from American Type Culture Collection (ATCC). The non‐tumor trophoblast HTR‐8/SVneo was a gift from Prof. Charles Graham (Queen's University at Kingston, Canada). Human umbilical vein endothelial cells (HUVECs; originally from ATCC) and aortic endothelial cells (HAoECs; originally from PromoCell GmbH) were provided by Dr. Andriana Margariti and Prof. David Grieve, respectively (Queen's University Belfast, UK). BeWo cells were maintained in DMEM/F12 Gibco (Thermo Fisher) supplemented with 2 mmol/L L‐glutamate. Jar and HTR‐8/SVneo were cultured in RPMI 1640 (Sigma‐Aldrich), and Jeg‐3 was cultured in EMEM (ATCC). Both HUVECs and HAoECs were cultured in EGM‐2 (Lonza) with all vendor‐provided supplements in flasks or plates coated with 0.1% gelatin (Sigma‐Aldrich). All growth media contained 10% fetal calf serum. There was no mycoplasma contamination with the cells.

Zhao J., Chow R.P., McLeese R.H., Hookham M.B., Lyons T.J, & Yu J.Y. (2020). Modelling preeclampsia: a comparative analysis of the common human trophoblast cell lines. FASEB BioAdvances, 3(1), 23-35.

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Pullulan-based Fluorescent Conjugates

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Pullulan (110 kDa, polydispersity index 2.25) and rhodamine B isothiocyanate (RhB-ITC) were purchased from Sigma-Aldrich (Milan, Italy). Salts, solvents and all the other chemical reagents were obtained from Carlo Erba (Milan, Italy), VWR (Milan, Italy) and Sigma-Aldrich (Milan, Italy). Azide-terminating amino-poly(ethylene glycol) (N 3 -PEG 350 -NH 2 ) and azide-terminating hydroxy-poly(ethylene glycol) (N 3 -PEG 395 -OH) were purchased from IRIS Biotech GmbH (Marktredwitz, Germany). DMEM-F12 Gibco® and TryPLE Express were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Foetal bovine serum (FBS), streptomycin, Lglutamine, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and all other products for cell culture were obtained from Sigma-Aldrich (Milan, Italy). 4′,6-diamidine-2′phenylindole dihydrochloride (DAPI) was obtained from Sigma-Aldrich (Milan, Italy) and wheat germ agglutinin-Alexa Fluor 633 (WGA-AlexaFluor 633) was obtained from Thermo Fisher Scientific (Madison, WI, USA).

Balasso A., Subrizi A., Salmaso S., Mastrotto F., Garofalo M., Tang M., Chen M., Xu H., Urtti A, & Caliceti P. (2021). Screening of chemical linkers for development of pullulan bioconjugates for intravitreal ocular applications. European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences, 161.

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Caco-2 Cell Inflammatory Model Protocol

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The normal intestinal epithelial cell line Caco-2 was purchased from American Type Culture Collection. The cells were cultured in Dulbecco's modified Eagle medium (DMEM)/F12 (Gibco; Thermo Fisher Scientific, Inc.) containing 10% fetal calf serum (FBS; HyClone; GE Healthcare Life Sciences) and 1% penicillin-streptomycin and were maintained at 37˚C with 5% CO₂ in an incubator. Caco-2 cells were treated with 2% DSS at 37˚C for 4 days to construct the inflammatory model (18 (link)).

Deng X., Shang L., Du M., Yuan L., Xiong L, & Xie X. (2020). Mechanism underlying the significant role of the miR-4262/SIRT1 axis in children with inflammatory bowel disease. Experimental and Therapeutic Medicine, 20(3), 2227-2235.

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Isolation and Culture of Mouse Tail Fibroblasts

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Using P2 mice, a 1–2 mm piece of tail was rinsed once in 70% ethanol and then twice in PBS containing 1X Antibiotic-Antimycotic (Gibco, 15240096). Next, tails were minced in 0.25 mL mixture of collagenase (Worthington Labs, LS004204, diluted in RPMI1640 plus L-glut, Gibco, 11875). After mincing, an additional 0.25 mL collagenase mixture was added, and the tissue was incubated for 30 minutes at 37°C. 6 mL MEF media (DMEM/F12 Gibco, 11330 + 10% FBS + PSG) was added, and tissues were incubated overnight at 37°C in 5% CO₂. Cells were incubated for an additional 3 days to allow adherence and spread away from remaining tissue. Tissue pieces were removed using sterile instruments, and cells were split and cultured as described above.

Schultz M.L., Schache K.J., Azaria R.D., Kuiper E.Q., Erwood S., Ivakine E.A., Farhat N.Y., Porter F.D., Pathmasiri K.C., Cologna S.M., Uhler M.D, & Lieberman A.P. (2022). Species-specific differences in NPC1 protein trafficking govern therapeutic response in Niemann-Pick type C disease. JCI Insight, 7(23), e160308.

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Molecular Characterization of Medulloblastoma Cell Lines

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Human MB cell lines D283 and UW-228 were originally obtained from the American Type Culture Collection (ATCC, Rockville, USA). These two cell lines present molecular features of different MB molecular subgroups: UW228 cells are TP53-mutated and classified as Shh, whereas D283 cells are p53 wild-type and classified as Group ¾ (Ivanov et al., 2016 (link)). The D283 cell line was cultured in Dulbecco’s modified Eagle’s medium (DMEM low glucose, Gibco, Grand Island, USA), while UW228 cell line was cultured in DMEM: Nutrient Mixture F-12 (DMEM/F-12 Gibco®), both medium supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco) and 1% (v/v) penicillin/streptomycin (Gibco). Cells were incubated in a humidified atmosphere of 5% CO₂ at 37°C.

Thomaz A., Pinheiro K.D., Souza B.K., Gregianin L., Brunetto A.L., Brunetto A.T., de Farias C.B., Jaeger M.D., Ramaswamy V., Nör C., Taylor M.D, & Roesler R. (2019). Antitumor Activities and Cellular Changes Induced by TrkB Inhibition in Medulloblastoma. Frontiers in Pharmacology, 10, 698.

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Cardiac Progenitor Cell Culture Protocols

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Explant culture: The anterior primitive streak (cardiac progenitor cells) was cultured in vitro for 4.5-Day as previously described [19] . Each treatment was performed in triplicate.
In the control group, approximately, 95% of the cells in the explant culture were determined to be cardiomyocytes which showed the presence of myosin heavy chain using MF20 immunofluorescent staining as previously reported [20] , and cell proliferation using pHIS3 immunofluorescent staining.
Primary culture: Primary cardiomyocyte cultures were established from day 14 chick embryo hearts.
Cell culture: H9c2 cells were purchased from Guangzhou Jennio Biotech Co., Ltd, China and cultured in culture medium (DMEM-F12 GIBCO).

Gao L.R., Wang G., Zhang J., Li S., Chuai M., Bao Y., Hocher B, & Yang X. (2018). High salt-induced excess reactive oxygen species production resulted in heart tube malformation during gastrulation. Journal of cellular physiology, 233(9).

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Breast Cancer Cell Line Zoledronate Study

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Human BC cell lines, MCF-7 and SkBr3, purchased from the American Type Culture Collection (Rockville, MD, USA) were grown in Dulbecco's modified Eagle's medium Gibco DMEM:F12 (Invitrogen, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS) and 1% Penicillin/Streptomycin (P/S) (Gibco). Cells were incubated at 37°C in a humidified atmosphere of 5% of CO₂. Eighty per cent confluent cultures were stimulated with ZOL 10 μM for 24 h. ZOL was kindly provided by Novartis Pharma AG.

Fanale D., Amodeo V., Bazan V., Insalaco L., Incorvaia L., Barraco N., Castiglia M., Rizzo S., Santini D., Giordano A., Castorina S, & Russo A. (2016). Can the microRNA expression profile help to identify novel targets for zoledronic acid in breast cancer?. Oncotarget, 7(20), 29321-29332.

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Parasite Infection Fluorescence Assay

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HFFs were grown on glass-bottom dark 24-well plates (Greiner Bio-One) and the confluent monolayers were infected with different parasite strains at an MOI of 1 in DMEM containing 1% FBS. At 24 h post-infection, the cells were washed with PBS after which Gibco DMEM/F-12 (Invitrogen) medium without phenol red and supplemented with 10 μM 5(6)-Carboxy-2’,7’-dichlorofluorescein diacetate (CDCFDA) was added to the cells. After 10 minutes of incubation at 37°C, the medium was removed and the cells were washed three times with PBS. Gibco DMEM/F-12 growth medium was added to the cells and the cells were imaged immediately.

Paredes-Santos T.C., Bitew M.A., Swale C., Rodriguez F., Krishnamurthy S., Wang Y., Maru P., Sangaré L.O, & Saeij J.P. (2023). Genome-wide CRISPR screen identifies genes synthetically lethal with GRA17, a nutrient channel encoding gene in Toxoplasma. PLOS Pathogens, 19(7), e1011543.

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