Dmem f12

Manufactured by Fujifilm

Sourced in Japan, United States

DMEM/F12 is a cell culture medium designed for the growth and maintenance of a wide variety of cell types. It is a balanced salt solution that provides essential nutrients, vitamins, and other components required for cell proliferation and survival. The medium is formulated to support the growth of both adherent and suspension cell lines.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

DMEM/F12 medium (13 citations) Advanced DMEM/F12 (2 citations) Dulbecco’s Modified Eagle’s Medium/Ham’s Nutrient Mixture F-12 (DMEM/Ham’s-F12) (2 citations)

68 protocols using dmem f12

Generating Trisomy 21 and Corrected iPSC Lines

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human iPSCs were cultured as previously described [2 (link), 22 (link)]. In brief, iPSCs were maintained on mitomycin C (Sigma)-inactivated mouse embryonic fibroblasts in human ES cell medium consisting of DMEM/F12 (Wako) supplemented with 20% KnockOut Serum Replacement (Gibco), 2 mM L-alanyl-L-glutamine (Wako), 1% MEM nonessential amino acid solution (Wako), 0.1 mM 2-mercaptoethanol (Sigma), and 5 ng/mL basic fibroblast growth factor (Katayama Chemical). Cultures were passaged every 6–8 days either manually or enzymatically using dispase II (Roche, Basel, Switzerland).
Secondary fibroblast-like cells were differentiated as described previously, with some modifications [23 (link), 24 (link)]. Briefly, embryoid bodies made from human iPSCs were cultured for 4 days in nonadherent cell culture plates in differentiation medium (80% knockout DMEM (KO-DMEM; Thermo Fisher Scientific Inc.), 1 mM L-glutamine, 0.1 mM β-mercaptoethanol, 20% FBS, and 1% nonessential amino acids.) Next, the cell aggregates were seeded into gelatin-coated plates and cultured for an additional 9 days. The outgrowing cell population was used as secondary fibroblast-like cells after at least two passages, and cultured in the medium described above. One clone each of a trisomy 21 fibroblast-derived human iPSC line (Tri21 iPSCs) and corrected disomy 21 iPSC line (cDi21 iPSC) was used in the experiments.

Nawa N., Hirata K., Kawatani K., Nambara T., Omori S., Banno K., Kokubu C., Takeda J., Nishimura K., Ohtaka M., Nakanishi M., Okuzaki D., Taniguchi H., Arahori H., Wada K., Kitabatake Y, & Ozono K. (2019). Elimination of protein aggregates prevents premature senescence in human trisomy 21 fibroblasts. PLoS ONE, 14(7), e0219592.

+ Open protocol

+ Expand

Chondrogenic Dedifferentiation Evaluation

Check if the same lab product or an alternative is used in the 5 most similar protocols

To evaluate whether dedifferentiation of ROCKi-treated cells was suppressed, immunohistochemical staining was performed to evaluate the regenerative ability within chondrogenic pellet culture. Monolayer-cultured cells up to P2 under treated and non-treated conditions were detached by trypsin, and pellet cultures were prepared by adding 2.5 × 10⁵ cells in 1 ml medium to 15 ml conical polypropylene tubes. The cells were pelleted by centrifugation at 400×g for 6min and subsequently cultured at 37°C, 21% O₂, and 5% CO₂. Pellets were maintained in 5% FBS, 5 ng/mL TGF-β1 (PeproTech, Rocky Hill, NJ), 10 ng/mL R3 IGF-1 (Abcam, UK), 5 mg/mL recombinant human insulin, 5 mg/mL human transferrin, and 5 ng/mL selenous acid (ITS-Premix, BD Bioscience) Dulbecco's modified Eagle's medium/nutrient mixture F-12 (DMEM:F12, Wako) as previously described¹⁹⁾ and were cultured for 2 weeks¹⁶⁾.

Nukaga T., Sakai D., Schol J., Suyama K., Nakai T., Hiyama A, & Watanabe M. (2019). Minimal Sustainability of Dedifferentiation by ROCK Inhibitor on Rat Nucleus Pulposus Cells In Vitro. Spine Surgery and Related Research, 3(4), 385-391.

+ Open protocol

+ Expand

Maintenance and Passaging of Human Embryonic Stem Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human ESCs (KhES-1) were used in accordance with ‘The Guidelines for Derivation and Utilization of Human Embryonic Stem Cells’ of the Ministry of Education, Culture, Sports, Science and Technology of Japan after approval by the Institutional Review Board. hESCs were maintained and cultured as previously described (Sakaguchi et al., 2015 (link)). In brief, hESCs were maintained on a feeder layer of mouse embryonic fibroblasts inactivated by mitomycin C treatment in DMEM/F12 (Wako) supplemented with 20% knockout serum replacement (KSR, Invitrogen), 2 mM glutamine, 0.1 mM nonessential amino acids (Invitrogen), 5 ng ml⁻¹ recombinant human basic fibroblast growth factor (bFGF) (Wako) and 0.1 mM 2-mercaptoethanol under 2% CO₂. For passaging, hESC colonies were detached and recovered en bloc from the feeder layer by treating them with 0.25% (weight/vol) trypsin and 1 mg ml⁻¹ collagenase IV in PBS containing 20% (vol/vol) KSR and 1 mM CaCl₂ at 37°C for 7-8 min. The detached hESC clumps were broken into smaller pieces (several dozens of cells) by gentle pipetting. The passages were performed at a 1:4 to 1:6 split ratio every 4-5 days.

Ogura T., Sakaguchi H., Miyamoto S, & Takahashi J. (2018). Three-dimensional induction of dorsal, intermediate and ventral spinal cord tissues from human pluripotent stem cells. Development (Cambridge, England), 145(16), dev162214.

+ Open protocol

+ Expand

Establishment and Characterization of Trophoblast Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

TS^mole cells were established as described previously (14 (link)). Briefly, CT cells were isolated from CHM tissues and cultured on plates coated with 5–10 µg/mL Col IV (Corning) using TS medium (DMEM/F12 [Wako] supplemented with 0.1 mM 2-mercaptoethanol [Wako], 0.2% FBS [Thermo Fisher Scientific], 0.5% Penicillin-Streptomycin [Thermo Fisher Scientific], 0.3% BSA [Wako], 1% ITS-X supplement [Wako], 1.5 μg/mL l-ascorbic acid [Wako], 50 ng/mL EGF [Wako], 2 μM CHIR99021 [Wako], 0.5 μM A83-01 [Wako], 1 μM SB431542 [Wako], 0.8 mM VPA [Wako], and 5 μM Y27632 [Wako]). TS^bip #1, #2, and #3 were established in our previous study (14 (link)) and correspond to TS^CT #1, #2, and #3, respectively. TS^bip #4 was established in this study and used only for CNV analysis. Unless otherwise noted, we used TS^mole and TS^bip cells passaged 10–20 times for the analysis.

Takahashi S., Okae H., Kobayashi N., Kitamura A., Kumada K., Yaegashi N, & Arima T. (2019). Loss of p57KIP2 expression confers resistance to contact inhibition in human androgenetic trophoblast stem cells. Proceedings of the National Academy of Sciences of the United States of America, 116(52), 26606-26613.

+ Open protocol

+ Expand

Breast Cancer Cell Line Cultivation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Breast cancer cell lines MCF7 (AKR-211, MCF-7/GFP; Cell Biolabs, Inc., San Diego, CA, USA) and MDA-MB231 (AKR-201, MDA-MB231/GFP; Cell Biolabs, Inc., San Diego, CA, USA) were cultivated in DMEM/F12 (Wako Pure Chemical, Osaka, Japan) supplemented with 10% FBS (Origen Mexico; Gibco, Thermo Fisher Scientific, Waltham, MA, USA). The non-carcinogenic breast epithelial cell line MCF10A (CRL-10317; ATCC, Manassas, VA, USA) was cultivated on a collagen-coated dish in HuMEC (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C under 5% CO₂. For measurements, cells were cultured on collagen gel (Cellmatrix type I-A; Nitta Gelatin, Osaka, Japan) on a 14- or 22-mmφ cover glass (#1; Matsunami Glass Ind. Ltd., Osaka, Japan) for 1–4 days at 37 °C under 5% CO₂, in sub-confluent to confluent conditions. In some experiments, 3 types of cells (e.g., different cell lines or different shRNA treated cells) each on 3 or 6 separated collagen-gel patches were cultured simultaneously on a 22-mmφ cover glass, which made it easy to compare the responses under the same conditions of cultivation and measurement.

Furuya K., Hirata H., Kobayashi T, & Sokabe M. (2021). Sphingosine-1-Phosphate Induces ATP Release via Volume-Regulated Anion Channels in Breast Cell Lines. Life, 11(8), 851.

+ Open protocol

+ Expand

Sevoflurane Exposure in Glioblastoma Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

High-grade glioma patient-derived neurospheres (MD13 and Me83, which are known as human mesenchymal glioblastoma stem cells) were established from surgical specimens obtained by Dr. Nakano and his colleagues and molecularly characterized as previously described [6 (link)]. This study used cultured cells but did not use specific patient information. Glioblastoma stem cells (GSCs) were cultured in serum-free neurosphere medium [DMEM/F12 (048-29785; FUJIFILM Wako Pure Chemical, Osaka, Japan) supplemented with B27 (1:50, 130-097-263; Miltenyi Biotec, Bergisch Gladbach, Germany), penicillin/streptomycin (P4333; Sigma-Aldrich, St. Louis, MO, USA), L-glutamine (G7513, Sigma-Aldrich), basic fibroblast growth factor (20 ng/mL, 100-18B; PeproTech, Cranbury, NJ, USA), and epidermal growth factor (20 ng/mL; AF-100-15, PeproTech)] at 37 °C. Cells in the exponential growth phase were plated into 96-well ultralow plates (Corning, 3474) and treated with 2% sevoflurane (Maruishi Pharmaceutical, Osaka, Japan) in a sealed modular incubator chamber (MIC-101; Billups-Rothenburg, Del Mar, CA, USA). The cells in the sevoflurane groups were exposed to sevoflurane for 2, 4, and 6 h, and 6 days. Cells exposed to air were used as a control group.

Shoji T., Hayashi M., Sumi C., Kusunoki M., Uba T., Matsuo Y, & Hirota K. (2022). Sevoflurane Does Not Promote the Colony-Forming Ability of Human Mesenchymal Glioblastoma Stem Cells In Vitro. Medicina, 58(11), 1614.

+ Open protocol

+ Expand

Establishment of Immortalized Cell Lines

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

For cell line development, fresh xenograft tissues were prepared as previously described [34 (link)]. Cells were cultured in DMEM/F12 (Wako) containing 1–10% FBS and insulin-transferrin-selenium (ITS, Gibco BRL, Carlsbad, CA, USA). Stromal cells were sequentially removed by partial trypsinization and mechanical removal. Cancer cells were subsequently cultured in DMEM containing 10% FBS when becoming morphologically homogenous. All media were supplemented with 100 U/mL penicillin and 100 μg/mL streptomycin. The cultures were maintained at 37 °C in a humidified 5% CO₂ atmosphere. Cells were maintained in vitro culture system at least 6 months to ensure the immortalization properties.
All four newly established cell lines were deposited into the Japanese Cancer Research Resources Bank (JCRB), National Institutes of Biomedical Innovation, Health and Nutrition (NIBIOHN), Osaka, Japan.

Vaeteewoottacharn K., Pairojkul C., Kariya R., Muisuk K., Imtawil K., Chamgramol Y., Bhudhisawasdi V., Khuntikeo N., Pugkhem A., Saeseow O.T., Silsirivanit A., Wongkham C., Wongkham S, & Okada S. (2019). Establishment of Highly Transplantable Cholangiocarcinoma Cell Lines from a Patient-Derived Xenograft Mouse Model. Cells, 8(5), 496.

+ Open protocol

+ Expand

Human Embryonic Stem Cell Maintenance and Passage

Check if the same lab product or an alternative is used in the 5 most similar protocols

We used the human embryonic stem cell (hESC) line H9. These PSCs were maintained on mitomycin-C-treated SNL feeder cells in DMEM/F12 (FUJIFILM Wako Pure Chemical) hESC medium containing 20% KnockOut serum replacement (Life Technologies), 1% nonessential amino acids (Sigma), 2 mM L-glutamine (Nacalai Tesque), 0.1 mM 2-mercaptoethanol (Sigma), 4 ng/mL FGF2 (PeproTech), 100 U/mL penicillin and 100 μg/mL streptomycin. Cells were passaged onto feeder cells or into feeder-free culture using a dissociation solution containing 0.25% trypsin, 100 mg/mL collagenase type IV (Invitrogen), 1 mM CaCl₂ and 20% KnockOut serum replacement (KSR). For feeder-free culture, cells were passaged on Matrigel (Corning, #354277)-coated dishes in SNL feeder-conditioned hESC medium containing 5 ng/mL FGF2.

Saeki T., Yoshimatsu S., Ishikawa M., Hon C.C., Koya I., Shibata S., Hosoya M., Saegusa C., Ogawa K., Shin J.W., Fujioka M, & Okano H. (2022). Critical roles of FGF, RA, and WNT signalling in the development of the human otic placode and subsequent lineages in a dish. Regenerative Therapy, 20, 165-186.

+ Open protocol

+ Expand

Isolation of Rat Alveolar Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fisher 344 rats (3–4 weeks old) were exposed to general anesthesia. The pulmonary artery was cannulated and perfused with 25 mL PBS containing 50 U/mL heparin (Mochida, Tokyo, Japan) and 1 μg/mL sodium nitroprusside (Sigma-Aldrich), and the lungs were removed. The alveoli were washed twice by injecting and draining PBS from the trachea. The alveoli were then filled with solution A (DMEM/F-12 + 2.5% HEPES (Wako) + 4.5 U/mL elastase (Worthington, Lakewood, NJ, USA) + 0.02 mg/mL DNase I (Sigma-Aldrich)), placed in a flask containing solution A, and shaken in a shaker at 37 °C for 45 min. The peripheral two-thirds of the lungs were then minced, placed in a new flask with solution A, and shaken for 15 min in a shaker at 37 °C. After shaking, a solution containing FBS (DMEM/F-12 + 2.5% HEPES + 50% FBS) was added, and the reaction was stopped by cooling on ice for 5 min. After successive filtration through 100- and 70-µm cell strainers, the lung cells were separated by centrifugation (300× g, 5 min), and the resulting cell pellets were resuspended in DMEM/F-12.

Ishii M., Tsuchiya T., Doi R., Morofuji Y., Fujimoto T., Muto H., Suematsu T., Mori R., Matsumoto K., Miyazaki T., Tomoshige K., Watanabe H., Iwatake M, & Nagayasu T. (2021). Increased In Vitro Intercellular Barrier Function of Lung Epithelial Cells Using Adipose-Derived Mesenchymal Stem/Stromal Cells. Pharmaceutics, 13(8), 1264.

+ Open protocol

+ Expand

Human iPS Cell Culture Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human iPS cell line Windy was provided by Dr. Akihiro Umezawa, National Center for Child Health and Development. Cells were cultured on mitomycin C-treated mouse embryonic fibroblasts in a 1:1 mixture of Dulbecco’s modified Eagle’s medium (DMEM) and Ham’s nutrient mixture F-12 (DMEM/F-12) (Wako Pure Chemical Industries, Osaka, Japan) basal medium with 20% KnockOut Serum Replacement (Thermo Fisher Scientific, Carlsbad, CA, USA), 2 mM l-glutamine (Wako Pure Chemical Industries), 1% minimum essential medium nonessential amino acid solution (NEAA) (Wako Pure Chemical Industries), 0.1 mM 2-mercaptoethanol (β-MeE) (Sigma-Aldrich, St. Louis, MO, USA), and 5 ng/mL basic fibroblast growth factor (FGF2) (PeproTech Inc., Rocky Hill, NJ, USA).

Qiu S., Kabeya T., Ogawa I., Anno S., Hayashi H., Kanaki T., Hashita T., Iwao T, & Matsunaga T. (2020). Gellan Gum Promotes the Differentiation of Enterocytes from Human Induced Pluripotent Stem Cells. Pharmaceutics, 12(10), 951.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!