Mem non essential amino acids solution

Manufactured by Merck Group

Sourced in United Kingdom, United States, Germany, Switzerland

MEM non-essential amino acids solution is a lab equipment product manufactured by Merck Group. It is a ready-to-use solution containing a mixture of non-essential amino acids commonly used in cell culture media and applications that require a standardized source of non-essential amino acids.

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MEM non-essential amino acids (127 citations) MEM non-essential amino acid solution (96 citations) Amino acids (83 citations) MEM amino acids solution (11 citations) MEM amino acids (10 citations) Essential amino acids (9 citations) MEM amino acid solution (7 citations) Amino acids solution (3 citations)

12 protocols using mem non essential amino acids solution

Embryo Culture Media Optimization

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Flush medium: CMRL 1066 (11 530 037, Invitrogen) + 5 × penicillin‐streptomycin (60162ES76, YEASEN) + 10% FBS (SE200‐ES, Vistech).
Medium I for embryos on IVC days 0–1: CMRL 1066+ 1 × penicillin‐streptomycin+ 1 × GlutaMAX Supplement (35 050 061, Thermo) + 1 × MEM Non‐Essential Amino Acids Solution (11 140 050, Thermo) + 0.5 × N‐2 Supplement (17 502 048, Gibco) + 0.5 × B‐27 Supplement (17 504 044, Gibco) + 10% FBS.
Medium II for embryos on IVC day 2: CMRL 1066 + 1 × penicillin‐streptomycin + 1 × GlutaMAX Supplement+ 1 × MEM Non‐Essential Amino Acids Solution + 0.5 × N‐2 Supplement + 0.5 × B‐27 Supplement + 20% FBS.
Medium III for embryos on IVC days 3–4: CMRL 1066 + 1 × penicillin‐streptomycin + 1 × GlutaMAX Supplement + 1 × MEM Non‐Essential Amino Acids Solution + 30% KnockOut Serum Replacement (10 828 028, Gibco).
Medium IV for embryos on IVC day 5: CMRL 1066 + 1 × penicillin‐streptomycin+ 1 × GlutaMAX Supplement + 1 × MEM Non‐Essential Amino Acids Solution + 30% KnockOut Serum Replacement + 30% RS (rat serum) + 0.5 mg mL⁻¹ glucose (Sigma, D9434).
Medium V medium for embryos on IVC days 6–10: CMRL 1066+ 1 × penicillin‐streptomycin + 1 × GlutaMAX Supplement + 1 × MEM Non‐Essential Amino Acids Solution + 50% RS + 1 mg mL⁻¹ glucose

Gu Z., Guo J., Zhai J., Feng G., Wang X., Gao Z., Li K., Ji S., Wang L., Xu Y., Chen X., Wang Y., Guo S., Yang M., Li L., Han H., Jiang L., Wen Y., Wang L., Hao J., Li W., Wang S., Wang H, & Gu Q. (2022). A Uterus‐Inspired Niche Drives Blastocyst Development to the Early Organogenesis. Advanced Science, 9(28), 2202282.

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Recellularization and Perfusion of Liver Scaffolds

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Livers recellularized with rat cells were perfused with Williams’ Medium E (Sigma) containing 2 mM GlutaMax-1 (Life Technologies), 0.1 mM MEM non-essential amino acids solution (Sigma), 10% fetal bovine serum (Hyclone), 10 μg/mL insulin (Sigma), 1 X penicillin/streptomycin (Life Technologies), and 1 μM dexamethasone (Sigma).
Scaffolds seeded with human liver cells were perfused with Williams’ Medium E (Sigma) containing 2 mM GlutaMax-1 (Life Technologies), 0.1 mM MEM nonessential amino acids solution (Sigma), 1X ITS+1 (Sigma), 2.0 ng/mL HGF (Peprotech), 2.0 ng/mL epidermal growth factor (EGF) (Peprotech), 1 X penicillin/streptomycin (Life Technologies), and 0.1 μM dexamethasone (Sigma).

Robertson M.J., Soibam B., O’Leary J.G., Sampaio L.C, & Taylor D.A. (2018). Recellularization of rat liver: An in vitro model for assessing human drug metabolism and liver biology. PLoS ONE, 13(1), e0191892.

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Expansion of Human T Cells from PBMCs

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Freshly collected PBMCs from the five donors were plated in 48-well plates (1 million cells/well) containing T cell media: RPMI-1640 medium (Gibco) with 5% human serum (Sigma-Aldrich), 1% Pen-Strep (Thermo Fisher Scientific), 1X MEM non-essential amino acids solution (Sigma-Aldrich), 1X sodium pyruvate (Gibco), and 1X HEPES buffer (Sigma-Aldrich), in addition to 1.6μL of Phytohemagglutinin-L (PHA-L) per well (500X, Invitrogen; Cat. no.: 00-4977-93) and 20 ng/mL of IL2 (BioLegend), and placed in a 5% CO₂ incubator at 37 °C. Cells were split every 2–3 days depending on density, and IL2 was replenished at a concentration of 20 ng/mL on the first two splits, then downscaled to 10 ng/mL. Cells were expanded for 12 days and were validated by flow cytometry using CD3-PE, CD4-PE and CD8-PB antibodies (Supplementary Fig. S6).

El Hajj Y., Shahin T., Dieng M.M., Alshaikh M., Khair M., Manikandan V, & Idaghdour Y. (2024). Pregnenolone sulfate induces transcriptional and immunoregulatory effects on T cells. Scientific Reports, 14, 6782.

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Preparation of Amino Acid-Deficient Media

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Amino acid-free (AA−) medium was prepared by RPMI 1640 powder (R8999–04A, US Biological Life Science) and sodium phosphate dibasic (5.6 mM, the same concentration as commercially available RPMI 1640 medium, US Biological Life Science), supplemented with 10% (v/v) dialyzed FBS (Thermo Fisher Scientific). Amino acid-sufficient (AA+) medium was prepared by adding proper volumes of MEM amino acids solution (essential amino acids, EAA, 50×), MEM non-essential amino acids solution (NEAA, 100×), and 200 mM L-Gln (all from Sigma-Aldrich) to AA− medium to reach a final concentration of 1×EAA, 1×NEAA, and 2 mM Gln. The medium was supplemented with 10% (v/v) dialyzed FBS. Medium containing single amino acids (Ala, Leu, Gln, or Arg) or their combinations was prepared with AA– medium (prepared to the same concentrations present in the AA+ medium). All media were adjusted to pH7.5 and filter-sterilized (0.2 μm) before use.

Zhu X., Wu Y., Li Y., Zhou X., Watzlawik J.O., Chen Y.M., Raybuck A.L., Billadeau D., Shapiro V., Springer W., Sun J., Boothby M.R, & Zeng H. (2024). Rag-GTPase-TFEB/TFE3 axis controls B cell mitochondrial fitness and humoral immunity independent of mTORC1. Research Square.

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Culturing Breast Cancer Cell Lines

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4T1-Luc cells were a generous gift from Prof. G. Lazennec (Centre national de la recherche scientifique (CNRS), Montpellier, France). Cells were cultured in DMEM’s media (Sigma-Aldrich, Dorset, UK) supplemented with 10% FBS, 5 mL of MEM Non-Essential Amino Acids Solution (100×), 100 U/mL penicillin, 100 μg/mL streptomycin, 0.146 g/L l-glutamine (Sigma-Aldrich) and G418 (500 µg/mL).
Cells were split the day prior to injection and harvested at around 40–50% confluency.
MDA-MB-231 and MCF-7 were cultured in Dulbecco’s Modified Eagle Media (DMEM) without phenol red (Sigma-Aldrich) supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 0.146 g/L l-glutamine (Sigma-Aldrich). Primary human mammary epithelial cells (pHMEC) were cultured in HuMEC basal serum free media (Thermofisher Scientific, Paisley, UK).

del Molino del Barrio I., Meeson A., Cooke K., Malki M.I., Barron-Millar B., Kirby J.A, & Ali S. (2021). Contribution of Heparan Sulphate Binding in CCL21-Mediated Migration of Breast Cancer Cells. Cancers, 13(14), 3462.

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Culturing and Differentiating Mouse Embryonic Stem Cells

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The HEK (Human Embryonic Kidney) 293T-17 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Hyclone, South Logan, UT, USA) with 10% fetal bovine serum (FBS; Hyclone), 50 units/mL penicillin (Sigma Aldrich, Saint Louis, MO, USA), and 50 μg/mL streptomycin (Sigma Aldrich) at 37 °C with 5% CO₂. We cultured the mESC lines CCE (129/sv) and J1 (129S4/SvJae) under feeder-free conditions and maintained in DMEM with 15% heat-inactivated FBS, 1X MEM Non-Essential Amino Acids Solution (Sigma Aldrich), 300 μM monothioglycerol (Sigma Aldrich), 1X penicillin/streptomycin, and 1000 U/mL LIF (leukemia inhibitory factor, Sigma Aldrich) in 0.1% gelatin-coated cell culture dishes at 37 °C with 5% CO₂. For differentiation, a hanging drop method [34 (link)] was employed. Cell aggregates were transferred to bacteria culture dishes on day 2 and further cultured in ESC medium without LIF. We monitored stemness by alkaline phosphatase staining with the Leukocyte Alkaline Phosphatase Assay Kit (Sigma Aldrich).

Lee Y.J., Son S.H., Lim C.S., Kim M.Y., Lee S.W., Lee S., Jeon J., Ha D.H., Jung N.R., Han S.Y., Do B.R., Na I., Uversky V.N, & Kim C.G. (2020). MMTR/Dmap1 Sets the Stage for Early Lineage Commitment of Embryonic Stem Cells by Crosstalk with PcG Proteins. Cells, 9(5), 1190.

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Amino Acid Modulation of Cell Culture Medium

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Amino acid-free (AA−) medium was prepared by RPMI 1640 powder (R8999–04A, US Biological Life Science) and sodium phosphate dibasic (5.6 mM, the same concentration as commercially available RPMI 1640 medium, US Biological Life Science), supplemented with 10% (v/v) dialyzed FBS (Thermo Fisher Scientific). Amino acid-sufficient (AA+) medium was prepared by adding proper volumes of MEM amino acids solution (essential amino acids, EAA, 50×), MEM non-essential amino acids solution (NEAA, 100×), and 200 mM L-Gln (all from Sigma-Aldrich) to AA- medium to reach a final concentration of 1×EAA, 1×NEAA, and 2 mM Gln. The medium was supplemented with 10% (v/v) dialyzed FBS. Medium containing single amino acids (Ala, Leu, Gln, or Arg) or their combinations was prepared with AA− medium (prepared to the same concentrations present in the AA+ medium). All media were adjusted to pH7.5 and filter-sterilized (0.2 μm) before use.

Zhu X., Wu Y., Li Y., Zhou X., Watzlawik J.O., Chen Y.M., Raybuck A.L., Billadeau D., Shapiro V., Springer W., Sun J., Boothby M.R, & Zeng H. (2024). The nutrient-sensing Rag-GTPase complex in B cells controls humoral immunity via TFEB/TFE3-dependent mitochondrial fitness. bioRxiv.

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Neuroepithelial Cell Differentiation from iPSCs

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We differentiated iPSCs into neuroepithelial cells using a previously described differentiation protocol (Corti et al., 2012 ). iPSCs were seeded in neuronal medium based on Dulbecco’s modified Eagle’s medium/F12 (Gibco, Invitrogen), supplemented with MEM nonessential amino acids solution, N2, and heparin (2 μg/ml; Sigma-Aldrich), thus promoting neuroepithelial differentiation.
For cell expansion and clonal culture, NSCs were plated in growth medium (NEP medium) containing FGF with or without epidermal growth factor, as previously reported (Corti et al., 2008 (link)).

Simone C., Nizzardo M., Rizzo F., Ruggieri M., Riboldi G., Salani S., Bucchia M., Bresolin N., Comi G.P, & Corti S. (2014). iPSC-Derived Neural Stem Cells Act via Kinase Inhibition to Exert Neuroprotective Effects in Spinal Muscular Atrophy with Respiratory Distress Type 1. Stem Cell Reports, 3(2), 297-311.

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Comprehensive Reagents for Cell Metabolism Research

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The chemical and biochemical reagents were obtained from the following sources: ascorbic acid, l-buthionine sulfoximine (BSO), cystine, and L-γ-glutamyl-p-nitroanilide (GPNA) were purchased from Wako Pure Chemicals (Osaka, Japan); oligomycin and 3PO were from Calbiochem-Merck (Darmstadt, Germany); antimycin A1, bis-2-(5-phenylacetamido-1,3,4-thiadiazol-2-yl)ethyl sulfide (BPTES), cilengitide, dimethyl-α-ketoglutarate (DM-αKG), d-glucose, Dulbecco's modified Eagle's medium (DMEM), dimethylsulfoxide, l-glutamine, MEM amino acids solution, MEM non-essential amino acids solution, MEM vitamin solution, N-acetyl-cysteine (NAC), poly 2-hydroxyethyl methacrylate (poly-HEMA), proteinase and phosphatase inhibitor cocktail, sulfasalazine, and 2-deoxy-d-glucose (2DG) were from Sigma-Aldrich (St. Louis, MO, USA); CB839 and UK5099 were from Cayman Chemical (Ann Arbor, MI, USA); etomoxir was from Selleck Chemical (Houston, TX, USA). Antibodies against AMPKα, phosphorylated (p)-AMPKα (Thr172), ASCT2, FAK (Tyr397), p-FAK, IDH1, LKB1, PARP, p-H2AX (Ser139), and xCT were obtained from Cell Signaling (Beverly, MA). Antibodies against GCLC, GCLM, and ME1 were from Abcam (Cambridge, UK). Antibodies against α-tubulin and β-actin were from Wako Pure Chemicals. Antibodies against lamin B1 and Nrf2 were from MBL (Nagoya, Japan) and GeneTex (San Antonio, TX, USA), respectively.

Endo H., Owada S., Inagaki Y., Shida Y, & Tatemichi M. (2020). Metabolic reprogramming sustains cancer cell survival following extracellular matrix detachment. Redox Biology, 36, 101643.

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Generation and Maintenance of hiPSCs

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WT- and NS-iPSCs were previously generated from their respective dermal fibroblasts [24 (link)]. hiPSCs were maintained in hiPSC medium on mitomycin C (AG Scientific, San Diego, CA, USA)-treated mouse embryonic fibroblasts (feeder). The hiPSC medium consisted of DMEM/F-12 (Gibco, Thermo Fisher, Waltham, MA, USA) supplemented with 20% KnockOut™ serum replacement (Gibco), 1% penicillin-streptomycin (Gibco), 1% MEM non-essential amino acids solution, 0.1 mM β-mercaptoethanol (Sigma-Aldrich, Merck, Darmstadt, Germany), 1.2 mg/mL sodium carbohydrate (Sigma-Aldrich, St. Louis, MO, USA), and 10 ng/mL recombinant human bFGF (R&D Systems, Minneapolis, MN, USA). The hiPSCs were incubated at 37 °C in an atmosphere containing 5% CO₂ with a daily medium change, and passaged every 6 days.

Kim B., Koh Y., Do H., Ju Y., Choi J.B., Cho G., Yoo H.W., Lee B.H., Han J., Park J.E, & Han Y.M. (2022). Aberrant Cortical Layer Development of Brain Organoids Derived from Noonan Syndrome-iPSCs. International Journal of Molecular Sciences, 23(22), 13861.

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