Memα medium

Manufactured by Thermo Fisher Scientific

Sourced in United States

MEMα medium is a basal medium formulated for the cultivation of a variety of mammalian cell types. It provides the essential nutrients required for cell growth and maintenance in in vitro cell culture applications.

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Lab products found in correlation

41 protocols using memα medium

Isolation and Purification of Human Fetal Germ Cells

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Male gonads were dissected in saline solution (0.9% NaCl, Fresenius Kabi, Bad Homburg, Germany) and either fixed overnight in 4% (v/v) paraformaldehyde (PFA) for immunohistochemistry or disaggregated by enzymatic digestion, as previously described [30 (link)]. Briefly, male gonads were disaggregated with 1 mg/mL collagenase type IV (LS004186, CellSystems, Troisdorf, Germany) and 27 KUnits/mL RNase-free DNAse I (79254, Qiagen, Hilden, Germany) at 37 °C for 105 min (min). The cell suspension was centrifuged (1500 rpm) for 5 min, and the cells were resuspended in Minimum Essential Media (MEM) α medium (22561, ThermoFisher Scientific, Waltham, MA, United States) and plated in 0.1% (w/v) gelatin (G7041, Merck, Kenilworth, NJ, United States)-coated petri dish (10 cm diameter) (Corning, Corning, NY, United States) at 37 °C for 30 min in order to reduce the amount of gonadal somatic cells by differential cell adhesion selection. After incubation, the cell suspension enriched for hFGCs (non-adherent fraction) was collected for fluorescence-activated cell sorting (FACS) analysis or for further culture.

Martin-Inaraja M., Ferreira M., Taelman J., Eguizabal C, & Chuva De Sousa Lopes S.M. (2021). Improving In Vitro Culture of Human Male Fetal Germ Cells. Cells, 10(8), 2033.

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Expansion and Transduction of CD34+ Cells

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CD34⁺ cells were cultured 24 h at 1 × 10⁶ cells/ml in MEM-α medium (Thermo Fisher Scientific) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 1% penicillin/streptomycin and 2mM l-glutamine (Thermo Fisher), SCF (50 ng/ml), interleukin-3 (IL-3, 10 ng/ml), IL-6 (10 ng/ml), thrombopoietin (TPO, 10 ng/ml), Fms-like tyrosine kinase 3 (50 ng/ml), in a 37 °C incubator with a humidified atmosphere of 5% CO₂ in air. Cells were transduced at a multiplicity of infection (MOI) of 35–75 either with pRRL-EF1hsa-miR-150 or pRRL-H1-anti-miR-150 or pRRL-H1-shCTRL, sorted again 48 h after transduction using a cell sorter and anti-CD34 antibodies and cultured during 5 to 10 days in complete MEM-α with SCF (50 ng/ml) and human M-CSF (100 ng/ml). Only transduced-GFP⁺ cells were analyzed.

Selimoglu-Buet D., Rivière J., Ghamlouch H., Bencheikh L., Lacout C., Morabito M., Diop M., Meurice G., Breckler M., Chauveau A., Debord C., Debeurme F., Itzykson R., Chapuis N., Willekens C., Wagner-Ballon O., Bernard O.A., Droin N, & Solary E. (2018). A miR-150/TET3 pathway regulates the generation of mouse and human non-classical monocyte subset. Nature Communications, 9, 5455.

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Isolation and Culture of NK Cells

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Peripheral blood mononuclear cells (PBMCs) from 10 healthy volunteers and 10 HNSCC patients were isolated according to the manufacturer's protocol of the Ficoll‐Paque Premium kit (GE healthcare; VWR), and NK cells were isolated using a Miltenyi NK cell isolation kit (LS) (Miltenyi Biotech) following manufacturers’ procedures. Purity of the NK cells exceeded 85% and contained less than 1% CD3⁺ T cells. Isolated NK cells were cultured in a human NK cell culture medium with R10 + 5% human AB serum (Sigma).
The NK92 cell line was purchased from cell bank of China Academy of Sciences and was grown in a Minimum Essential Medium (MEM) α medium (Thermo Fisher Scientific) supplemented with 10% foetal bovine serum (FBS) (Thermo Fisher Scientific), 1.5 g/L NaHCO₃, 2 mM L‐glutamine, 0.2 mM inositol, 0.02 mM folic acid, 0.1 mM 2‐mercaptoethanol and 100 U/ml recombinant human interleukin‐2 (IL‐2). Cells were cultured in a 37℃ incubator under standard conditions with 5% CO₂.

Wang H., Nan S., Wang Y, & Xu C. (2021). CDX2 enhances natural killer cell–mediated immunotherapy against head and neck squamous cell carcinoma through up‐regulating CXCL14. Journal of Cellular and Molecular Medicine, 25(10), 4596-4607.

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Probiotic VSL#3 Impacts Inflammatory Pathways

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Sixty SD rats, aged 6–7 weeks and weighing 200–250 g, were separately fed in cages (5 rats for one cage). The rats were purchased from Better Biotechnology Co., Ltd., Nanjing, with an Item No. of J001; probiotic VSL#3 from Ferring Pharmaceuticals, USA; TNF-α ELISA kit from Tecan (Shanghai) Trading Co., Ltd., with an Item No. of BE45471; NF-κB Antibody from 51BioMart (Tianjin) Biotechnology Co., Ltd., with an Item No. of EL803353-100; TLR4 Antibody from Xiamen Huijia Biotechnology Co., Ltd., with an Item No. of IMG-6285A; β-Actin and horseradish peroxidase (HRP)-labeled goat anti-rat IgG (secondary antibody) from ABCAM, with Item No. of ab124964 and ab20272; fetal bovine serum (FBS), MEMα medium (low sugar), DMEM medium, Opti-MEM medium, ECL chemiluminescence detection kit, LipofectAMINE 2000 kit, RIPA and BCA protein kit from Thermo Fisher, Shanghai, China, with Item No. of 10099141, 12561072, 10569044, 11058021, 35050, 11668019, 23225 and 15224041; total RNA extraction kit EasyPure RNA Kit, PCR kit TransScript miRNA First-Strand cDNA Synthesis SuperMix and TransScript II Green Two-Step qRT-PCR SuperMix from TransGen Biotech, Beijing, China, with Item No. of ER101-01, AT351-01 and AQ301-01; 7500 PCR instrument from ABI, USA.
This experiment has been approved by the Animal Ethics Committee of our hospital.

WANG H., LI S., LI H., DU F., GUAN J, & WU Y. (2019). Mechanism of Probiotic VSL#3 Inhibiting NF-κB and TNF-α on Colitis through TLR4-NF-κB Signal Pathway. Iranian Journal of Public Health, 48(7), 1292-1300.

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Characterization of Leukemia Cell Lines

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MOLT3 and Jurkat cell lines were purchased from ATCC (Manassas, VA, USA); DND 41 and TALL1 cell lines were kindly provided by academic colleagues; SUPT11 cell lines were purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany); all these cell lines were cultured in complete RPMI medium, as reported elsewhere [33 (link)]. Cells were periodically tested for mycoplasma contamination. PDX cells were cultured in MEMα medium (Thermo Fisher Scientific) supplemented with 10% human heat inactivated AB⁺ serum, 10% fetal calf serum (FCS), 1% Glutamax, 1% penicillin/streptomycin and with 20 ng/ml FLT3 ligand, 10 ng/ml IL7, 50 ng/ml SCF (Peprotech, Rocky Hill, NJ, USA), and 20 nM human insulin (Sigma Aldrich, Saint Luis, MO, USA).
The following drugs were used: 0.5 µM TSA (Sigma Aldrich), 500 µM cyclohexamide (Sigma Aldrich), 20 µM MG132 (Sigma Aldrich), 20 µM CHL (Sigma Aldrich), 2 µM tubacin (Enzo Life Science, Farmingdale, NY), 100 nM bafilomycin (Sigma Aldrich), 2 µM HDAC1i and HDAC8i (Italfarmaco, Milan, Italy), 20 µM Ciliobrevin D. At planned time points, cells were harvested and processed for assessment of cell viability, caspase assay, and RNA and protein extraction.

Pinazza M., Ghisi M., Minuzzo S., Agnusdei V., Fossati G., Ciminale V., Pezzè L., Ciribilli Y., Pilotto G., Venturoli C., Amadori A, & Indraccolo S. (2018). Histone deacetylase 6 controls Notch3 trafficking and degradation in T-cell acute lymphoblastic leukemia cells. Oncogene, 37(28), 3839-3851.

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Isolation and Culture of Gingival Fibroblasts

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Small pieces of gingival tissue from freshly extracted third molar were collected and washed by saline solution containing 100 µg/mL penicillin/streptomycin (Thermo Fisher Scientific, MA, USA). Gingival tissue was cut and minced to 1.0 mm³, then incubated in digesting mixture containing 8 mg/mL neutral proteases and 6 mg/mL collagenase for 1 hr at 37°C. Gingival cells suspension was added in Minimum essential medium-α (MEM-α) medium (Thermo Fisher Scientific, MA, USA) consisting of 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, MA, USA) and 100 µg/mL penicillin/streptomycin (Thermo Fisher Scientific, MA, USA). Gingival fibroblasts were harvested and amplified by single cell colony methods. Cell culture was maintained at 37°C with 5% CO₂ in a humidified incubator before the following experiments.

Deng Z., Yu L., Kuang Y., Zhou Z, & Li X. (2024). Highly Ordered Nanotube-Like Microstructure on Titanium Dental Implant Surface Fabricated via Anodization Enhanced Cell Adhesion and Migration of Human Gingival Fibroblasts. International Journal of Nanomedicine, 19, 2469-2485.

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HeLa Cell Culture Protocol

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Hela cells were purchased from and authenticated by Atlantic Type Culture Collection. Cells were cultured in phenol red-free MEMα medium (Thermo Fisher) supplemented with 10% charcoal-stripped foetal bovine serum. Cells were maintained under standard culture conditions.

Mays S.G., Stec J., Liu X., D’Agostino E.H., Whitby R.J, & Ortlund E.A. (2020). Enantiomer-specific activities of an LRH-1 and SF-1 dual agonist. Scientific Reports, 10, 22279.

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Culturing MLO-Y4 and HEK293T Cells

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MLO-Y4 cells were established from female mice and cultured as previously described (Kato et al., 1997 ). The cells were seeded on type I collagen-coated 6 well plates under MEMα medium (Thermo Fisher Scientific, 12571–063), 2.5% (v/v) Fetal Bovine Serum (Hyclone, SH30396.03,Lot AB217307), 2.5% (v/v) calf serum (Hyclone, SH30072.03, AAL11105), penicillin- streptomycin (P/S) 100 U/ml. Cells were treated with irisin or other reagents at 60% cell density. HEK293T cells were set up for experiments at 1 × 10⁵ cells per well in 6 well plate under DMEM (Thermo Fisher Scientific, MT10017CV), 10% (v/v) Fetal Bovine Serum (Gemini BioProducts), penicillin-streptomycin (P/S) 100 U/ml. All cell lines were maintained at 37°C in 5% CO²

Kim H., Wrann C.D., Jedrychowski M., Vidoni S., Kitase Y., Nagano K., Zhou C., Chou J., Parkman V.J., Novick S.J., Strutzenberg T.S., Pascal B.D., Le P.T., Brooks D.J., Roche A.M., Gerber K.K., Mattheis L., Chen W., Tu H., Bouxsein M.L., Griffin P.R., Baron R., Rosen C.J., Bonewald L.F, & Spiegelman B.M. (2018). Irisin Mediates Effects on Bone and Fat via αV Integrin Receptors. Cell, 175(7), 1756-1768.e17.

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Isolation and Stimulation of Human Macrophages

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Peripheral blood mononuclear cells were obtained from blood leukocyte preparations (purchased from the New York Blood Center) by density-gradient centrifugation with lymphoprep (Accurate Chemical, cat #: AN1001969) following a protocol approved by the Hospital for Special Surgery Intuitional Review Board. CD14+ monocytes were purified from fresh peripheral blood mononuclear cells with anti-CD14 magnetic beads (Miltenyi Biotec, cat #: 130-050-201) as recommended by the manufacturer. Macrophages were derived by culture of monocytes for 24 hours in MEM-α medium (Invitrogen, cat #: 12561-072) supplemented with 10% (vol/vol) FBS (Hyclone, cat #: SH30070.03), 10% (vol/vol) L-Glutamine (Invitrogen, cat #: 25030-081), 10% (vol/vol) Penicillin/Streptomycin (Invitrogen, cat #: 15070-063), and human macrophage colony-stimulating factor (M-CSF; 10ng/ml; Peprotech, cat#: 300-25). DMSO or IBET151 (500nM) was added thirty minutes prior to the addition of the indicated stimulus. The stimuli were recombinant human TNF-α (10ng/ml), LPS (10ng/ml for mRNA analysis and 50ng/mL for ChIP), recombinant IFN-β (20units/ml for mRNA analysis and 200 units/ml for ChIP), recombinant human IFN-γ (100 units/ml), recombinant human IL-4 (100ng/ml), or recombinant human IL-10 (50ng/ml). The samples were harvested at the indicated time points.

Chan C.H., Fang C., Yarilina A., Prinjha R.K., Qiao Y, & Ivashkiv L.B. (2014). BET Bromodomain Inhibition Suppresses Transcriptional Responses to Cytokine-Jak-STAT Signaling in a Gene-specific Manner in Human Monocytes. European journal of immunology, 45(1), 287-297.

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Comparison of CHO cell lines

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The CHO AA8 (HR-proficient) and irs1SF (HR-deficient) cells were grown in MEMα medium (Invitrogen^™, Carlsbad, CA) with 10% fetal bovine serum (FBS) (Invitrogen), L-glutamine (Fisher Scientific, Boston, MA), and penicillin/streptomycin at 37°C, 5% CO₂. CHO cell doubling time was measured over 72 h. For evaluation of cell cycle progression, CHO cells were harvested, fixed and stained with propidium iodide. Cells were processed and analyzed by flow cytometry as described previously (9 (link)).

Im M.M., Flanagan S.A., Ackroyd J.J, & Shewach D.S. (2015). Drug Metabolism and Homologous Recombination Repair in Radiosensitization with Gemcitabine. Radiation research, 183(1), 114-123.

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