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Es e14tg2a

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The ES-E14TG2a is a cell line derived from mouse embryonic stem cells. It is a widely used model system for the study of embryonic stem cell biology and differentiation.

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L0022, by Biowest (1 mentions) Lm001 05, by Welgene (1 mentions) Esg1107, by Merck Group (1 mentions) Dmem f12, by Thermo Fisher Scientific (1 mentions) Mco 18aic, by Sanyo (1 mentions)

5 protocols using es e14tg2a

1

Localized Wnt3a Activation in mESCs

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Mouse embryonic stem cell ES-E14TG2a from ATCC (CRL-1821) were cultured on 0.1% gelatin-coated plates in DMEM medium containing 15% Fetal Bovine Serum, 1% Penicillin/Streptomycin, 1% Glutamax, 0.1 mM 2-mercaptoethanol, 1% MEM Non-Essential Amino Acids, 1% Sodium Pyruvate, and 1,000 U/ml recombinant leukemia inhibitory factor (LIF).
Wnt3a coated beads were prepared as described88 (link). Localized Wnt3a beads with mESCs were engineered in the 2 cm plates following the previous method15 (link). To achieve one cell and one bead contact, mESCs were seeded at a low concentration before imaging or collecting. After seeding the cells and Wnt3a beads, samples were collected at around 12 h.
Sun Z., Tang Y., Zhang Y., Fang Y., Jia J., Zeng W, & Fang D. (2021). Joint single-cell multiomic analysis in Wnt3a induced asymmetric stem cell division. Nature Communications, 12, 5941.
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2

Comparative Culture of mESCs

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All the mESCs described in this study are derived from ES-E14TG2a (ATCC CRL-1821). All the ES-E14TG2a derived mESCs used in this study were generated through CRISPR/Cas9-mediated genome editing, and their genotypes were confirmed by both PCR and western blot analysis. All the mESCs used in this study were cultured on 0.5% gelatin-coated tissue culture plates in either the 15% FBS + Lif (leukemia inhibitory factor) (medium DMEM/F-12 supplemented with 15% FBS, 2 mM L-glutamine, 0.1 mM MEM NEAA, 1% penicillin–streptomycin, 0.1 mM β-mercaptoethanol, and 1000 U/mL mLIF) or the 2i + Lif medium (DMEM/F-12, 2% FBS, 2 mM L-glutamine, 0.1 mM MEM NEAA, 1% penicillin–streptomycin, 0.1 mM β-mercaptoethanol and 1000 U/mL mLIF, 1 × N2N27, 3 µM CHIR99021 and 1 µM PD0325901). All the cells were grown in tissue culture incubators with temperature at 37°C and 5% CO2.
Liu Q., Chen X., Novak M.K., Zhang S, & Hu W. (2021). Repressing Ago2 mRNA translation by Trim71 maintains pluripotency through inhibiting let-7 microRNAs. eLife, 10, e66288.
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3

Culturing Mouse Embryonic Stem Cells

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mESCs (ES-E14TG2a) were purchased from the American Type Culture Collection (USA). mESCs were cultured on mitomycin C-treated mouse embryonic fibroblasts (mEFs) at humidified culture incubator (37°C with 5% CO2). The growth medium was prepared as in previously described [10 ].
Lee J.H., Yoo Y.M., Lee B., Jeong S., Tran D.N, & Jeung E.B. (2021). Melatonin mitigates the adverse effect of hypoxia during myocardial differentiation in mouse embryonic stem cells. Journal of Veterinary Science, 22(4), e54.
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4

Murine R1 ESCs Differentiation

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Cell Culture Murine R1 ESCs (Sirt1 +/+ and Sirt1 À/À ; Han et al., 2008) and ES-E14TG2a (purchased from American Type Culture Collection [ATCC]) were grown as described in the Supplemental Experimental Procedures. EB formation and in vitro neuronal or germ cell differentiation were performed using similar protocols to those used in previous studies (Payer et al., 2006; West et al., 2009) .
Heo J., Lim J., Lee S., Jeong J., Kang H., Kim Y., Kang J.W., Yu H.Y., Jeong E.M., Kim K., Kucia M., Waigel S.J., Zacharias W., Chen Y., Kim I.G., Ratajczak M.Z, & Shin D.M. (2017). Sirt1 Regulates DNA Methylation and Differentiation Potential of Embryonic Stem Cells by Antagonizing Dnmt3l. Cell reports, 18(8).
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5

Culturing Mouse Embryonic Stem Cells

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ES-E14TG2a, a mouse embryonic stem cell line (mESCs), was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA) and cultured using the same method described elsewhere [29 (link)]. The composition of the growth medium for maintaining undifferentiated cells was as follows: DMEM/F-12 (1:1) (11320-033, Gibco, Grand Island, NY, USA), 10% FBS (16000-044, Gibco, Grand Island, NY, USA), non-essential amino acid (NEAA; 11140-050, Gibco, Grand Island, NY, USA), plasmocin prophylactic (Plas; ant-mpp, InvivoGen, San Diego, CA, USA; 50 μg/mL), 100 U/mL penicillin and 100 mg/mL streptomycin (P/S; L0022, Biowest, Riverside, MO, USA), 2-mercaptoethanol (21985-023, Gibco, Grand Island, NY, USA; 10−4 M), and mouse leukemia inhibitory factor (mLIF; ESG1107, Millipore, Darmstadt, Germany; 10 ng/mL). Mouse embryo fibroblasts (mEFs) were used as the feeder cells. The mEFs were obtained from E10.5 mouse embryos [30 (link)]. The cells were grown at 37 °C in a 5% CO2 humidified tissue culture incubator (MCO-18AIC, Sanyo, Osaka, Japan). The 3T3-L1 cells (Clone A31 and ATCC) for the general toxicity test were cultured in DMEM (LM 001-05, WELGENE, Gyeongsan-si, Gyeongsangbuk-do, South Korea) with 10% fetal bovine serum (FBS; S1480-500, Biowest, Nuaillé, France) and P/S.
Jeong S., Ahn C., Kwon J.S., Kim K, & Jeung E.B. (2023). Effects of Sodium Arsenite on the Myocardial Differentiation in Mouse Embryonic Bodies. Toxics, 11(2), 142.
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