Embryomax dmem

Manufactured by Merck Group

EmbryoMax DMEM is a powdered culture medium formulated for the in vitro cultivation of embryonic stem cells, embryos, and other cell types. It provides a defined, serum-free environment to support the growth and maintenance of these cells.

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Lab products found in correlation

L glutamine, by Thermo Fisher Scientific (4 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Nucleoside, by Merck Group (2 mentions) Leukemia inhibitory factor, by Merck Group (2 mentions) 2 mercaptoethanol, by Merck Group (2 mentions) Pd98059, by Merck Group (2 mentions) Embryomax mem non essential amino acids, by Merck Group (2 mentions) Mercaptoethanol, by Merck Group (1 mentions) Mem non essential amino acid stock, by Thermo Fisher Scientific (1 mentions) Es 006 b, by Merck Group (1 mentions) Puromycin, by Thermo Fisher Scientific (1 mentions) Accutase, by Thermo Fisher Scientific (1 mentions) Glutamax 1 supplement, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Biowest (1 mentions) Embryomax fetal bovine serum, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Venor gem mycoplasma detection kit, by Merck Group (1 mentions) Embryomax nucleosides, by Merck Group (1 mentions) Esgro leukemia inhibitory factor, by Merck Group (1 mentions) β mercapthoethanol, by Merck Group (1 mentions)

Spelling variants of this product

EmbryoMax (31 citations)

6 protocols using embryomax dmem

Derivation and Maintenance of Mnx1::GFP ESCs

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The Mnx1::GFP ESC line was derived from blastocysts by mating a transgenic Mnx1::GFP male mouse with a wildtype female mouse, and the blastocysts were cultured in KES medium [EmbryoMax DMEM (Millipore) containing 1x EmbryoMax MEM Non-essential Amino Acids (Millipore), 1x Nucleosides (Millipore), 2 mM L-Glutamine (Invitrogen), 1% Penicillin/Streptomycin (Invitrogen), 0.00072% 2-mercaptoethanol (Sigma), 0.01% leukemia inhibitory factor (Millipore), 3

μ

M GSK3β inhibitor CHIR99021 (Merck), and 50

μ

M PD98059 (Merck)]. The Mnx1::GFP mouse ESC line with Litchi KO was generated by CRISPR/Cas9 gene editing of Mnx1::GFP ESCs (see Method details). To maintain ESCs, cells were cultured on gamma-inactivated mouse primary embryonic fibroblasts with ESC culture medium [15% Fetal Bovine Serum (GIBCO), 1x EmbryoMax MEM Non-essential Amino Acids (Millipore), 1x Nucleosides (Millipore), 2 mM L-Glutamine (Invitrogen), 1% Penicillin/Streptomycin (Invitrogen), 0.00072% 2-mercaptoethanol (Sigma), and 1000 μL/mL LIF/ESGRO (Chemicon/Millipore) in EmbryoMax DMEM (Millipore)], with culture medium replaced each day.

Hsu H.C., Hsu S.P., Hsu F.Y., Chang M, & Chen J.A. (2024). LncRNA Litchi is a regulator for harmonizing maturity and resilient functionality in spinal motor neurons. iScience, 27(3), 109207.

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Generation of Cstf2Gt(IST10905E6)Tigm Mouse ESCs

Cited 1 time

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Cstf2^{Gt(IST10905E6)Tigm} cell line was obtained from Texas A&M Institute for Genomic Medicine (TIGM) and derived from mouse C57BL/6N-derived Lex3.13 ESC lines in which a gene-trap cassette was inserted between the first and second exons (Cstf2^{Gt(IST10905E6)Tigm}, [23 ]). Mouse embryonic stem cells (ESCs) were maintained on 0.1% gelatin-coated 10 cm dishes without feeder cells in Embryo Max DMEM (Millipore) supplemented with 15% ESC-qualified fetal bovine serum (Hyclone/Thermo), 2mM L-glutamine (Gibco), 0.1mM -mercaptoethanol (Sigma), 0.1 mM MEM non-essential amino acid stock (Gibco), and 10 ng/mL human leukemia inhibitory factor (LIF, InVitria). ESCs were grown at 37°C in a humidified incubator in 5% CO₂ and passaged every two days (~70–80% confluency).
XEN cells [9 (link)] were a gift from Ann C. Foley (Clemson University), and were cultured in 10 cm dishes in ESC complete media without LIF at 37°C in a humidified incubator in 5% CO₂. For cardiomyocyte differentiation, XEN complete media was obtained from 70% confluent cells and 0.2 micron-filtered.

Youngblood B.A, & MacDonald C.C. (2014). CstF-64 is Necessary for Endoderm Differentiation Resulting in Cardiomyocyte Defects. Stem cell research, 13(3 0 0), 413-421.

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Culturing Mouse Embryonic Stem Cells

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Male mouse ES cells were purchased from Millipore (Nanog GFP Reporter cell line, SCR089). All cells were cultured in tissue culture flasks or 8 well μ-Slides (ibidi, 80826), coated with gelatin (0.1%) (Millipore, ES-006-B). ES cells were passaged using accutase (Life Technologies, A11105-01). Basic medium (BM) consisted of Embryomax DMEM (Millipore, SLM-220-B), supplemented with 15% (v/v) Embryomax fetal bovine serum (FBS) (Millipore, ES-009-C), 1 mM sodium pyruvate (Millipore, TMS-005-C), 0.1 mM non-essential amino acids (Life Technologies, 11140), and 2 mM Glutamax-I Supplement (Life Technologies, 35050). ES cells were cultured in basic growth medium (BGM), consisting of BM supplemented with 0.1 mM βME (Life Technologies, 31350-010), 10 ng/ml LIF (Life Technologies, PMC4054) and 0.5 μg/ml puromycin (Life Technologies, A11138-03). Immortalized MEFs were kindly provided by Reinhard Fässler (Max Planck Institute of Biochemistry, Martinsried, Germany). MEFs were cultured in DMEM (Life Technologies, 21885) supplemented with 10% (v/v) FBS (Biowest, S181H).

Moshfegh C., Aires L., Kisielow M, & Vogel V. (2016). A gonogenic stimulated transition of mouse embryonic stem cells with enhanced control of diverse differentiation pathways. Scientific Reports, 6, 25104.

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Generation and Characterization of Transgenic ESCs

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All the animals were genotyped by qPCR (Table S5). The smallest sample size was used that would still give a significant difference in accordance with the 3Rs. No animals were involved in previously unrelated experimental procedures. Analyses of the influence of sex was not evaluated in this study. All of the live animals were maintained in specific-pathogen-free (SPF) animal facility, approved and overseen by IACUC Academia Sinica.
Moues ESC derivation and culture ESC lines were derived from blastocysts by mating the double transgenic Foxp1::GFP; Hb9::RFP females with SOD1 G93A males, and the blastocysts were cultured in KES medium [EmbryoMax DMEM (Millipore) containing 1X EmbryoMax MEM Non-essential Amino Acids (Millipore), 1x Nucleosides (Millipore), 2 mM L-Glutamine (Invitrogen), 1% Penicillin/Streptomycin (Invitrogen), 0.00072% 2-mercaptoethanol (Sigma), 0.01% leukemia inhibitory factor (Millipore), 3 mM GSK3b inhibitor CHIR99021 (Merck), and 50 mM PD98059 (Merck)]. Stable ESC lines carrying both Foxp1::GFP; Hb9::RFP with or without the SOD1 G93A transgene were selected for further characterization.

Tung Y.T., Peng K.C., Chen Y.C., Yen Y.P., Chang M., Thams S, & Chen J.A. (2019). Mir-17∼92 Confers Motor Neuron Subtype Differential Resistance to ALS-Associated Degeneration. Cell stem cell, 25(2).

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Embryoid Body Formation from mESCs

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The mESCs were collected and centrifuged at 1,200 rpm for 1 min (KOKUSAN), and then the cells were suspended in mESC medium. After counting the cells, they were diluted to 2.0 × 10⁶ cells/mL for the manual and inkjet-dispensing methods, and to 3.0 × 10⁶ cells/mL for the FACS dispensing method. Cells (2.0 × 10³) were dispensed using the three methods and plated on 10 wells of U-bottom 384-well plates (Sumitomo Bakelite, Tokyo, Japan) for every method to prepare EBs. The mESC media were changed every 2 d. After 5 d, the EBs were collected from every well and transferred to six-well plates, and the media were also changed with EmbryoMax DMEM (Merck). After 9 d, the EBs were collected for qPCR analyses.

Yumoto M., Hemmi N., Sato N., Kawashima Y., Arikawa K., Ide K., Hosokawa M., Seo M, & Takeyama H. (2020). Evaluation of the effects of cell-dispensing using an inkjet-based bioprinter on cell integrity by RNA-seq analysis. Scientific Reports, 10, 7158.

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Mouse Embryonic Stem Cell Culture

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Mouse embryonic stem cells (ESCs) were cultured in EmbryoMax DMEM (EMD Millipore) supplemented with 15% embryonic stem cell screened fetal bovine serum (HyClone), 2mM L-glutamine (Life Technologies), 1x non-essential amino acids EmbryoMax MEM (EMD Millipore), 1x EmbryoMax nucleosides (EMD Millipore), 0.1mM β-mercapthoethanol (Sigma-Aldrich), 100U/ml penicillin-streptomycin (Life Technologies), 1000U/ml ESGRO Leukemia inhibitory factor (EMD Millipore), 2.5μM GSK-3 inhibitor XVI (EMD Millipore) and 50nM FGF receptor antagonist PD173074 (Tocris). NIH3T3 cells were grown in DMEM (Invitrogen) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific), 2mM glutamine (Life Technologies), and 100U/ml penicillin-streptomycin (Life Technologies). All cells were determined to be negative for mycoplasma using the Venor GeM Mycoplasma detection kit (Sigma-Aldrich).

Jacko M., Weyn-Vanhentenryck S.M., Smerdon J.W., Yan R., Feng H., Williams D.J., Pai J., Xu K., Wichterle H, & Zhang C. (2018). Rbfox splicing factors promote neuronal maturation and axon initial segment assembly. Neuron, 97(4), 853-868.e6.

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