Mem medium

Manufactured by Thermo Fisher Scientific

Sourced in United States, China, Germany, Japan, Belgium, United Kingdom

MEM medium is a cell culture medium designed for the in vitro maintenance and growth of a variety of mammalian cell types. It provides the necessary nutrients and growth factors to support cell proliferation and survival.

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Opti-MEM reduced serum medium (819 citations) MEM alpha medium (128 citations) Opti-MEMTM medium (13 citations) Advanced MEM medium (11 citations) Opti-MEMTM reduced serum medium (9 citations) Opti-MEMTM I Reduced Serum Medium (7 citations) Gibco® Opti-MEM™ medium (5 citations) Opti-MEM reduced serum-free medium (4 citations) Opti-Mem® I (1x) Reduced Serum Medium (3 citations) MEM glucose medium (3 citations)

359 protocols using mem medium

Cell Viability Assay of Fibroblasts

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Human fibroblast cells, MRC-5, were plated on 96-well flat bottom plates at a density of 1.5 × 10 4 cells/well with 100 µl of MEM medium (Life Technologies, Grand Islands, NY, USA) containing 10% fetal bovine serum (Hana-nesco Bio, Tokyo, Japan) and 1% penicillin-streptomycin (Life Technologies) and incubated at 37°C with 5% CO 2 for 48 h. Approximately 100 µl of MEM medium, containing 5 µl of each compound dissolved in 50% (v/v) aqueous DMSO, was added to each well. After 48 h cultivation at 37°C with 5% CO 2 , cell density and morphological changes were observed under a microscope. After observation, 10 µl of WST-8 solution (Dojindo, Kumamoto, Japan) was added to the cells and the plate was incubated at 37°C with 5% CO 2 for 2 h. The absorbance was measured at 450 nm by spectrophotometer (SH-9000 Lab., Corona Electric, Ibaraki, Japan). The growth of MRC-5 cells was measured in three independent experiments.

Pramisandi A., Dobashi K., Mori M., Nonaka K., Matsumoto A., Tokiwa T., Higo M., K.r.i.s.t.i.n.i.n.g.r.u.m., Amalia E., Nurkanto A., Inaoka D.K., Waluyo D., Kita K., Nozaki T., Ōmura S, & Shiomi K. (2020). Microbial inhibitors active against Plasmodium falciparum dihydroorotate dehydrogenase derived from an Indonesian soil fungus, Talaromyces pinophilus BioMCC-f.T.3979. The Journal of general and applied microbiology, 66(5).

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Culturing Human Breast Cancer Cell Lines

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Human breast cancer cell lines were used in this study. The BT-474 cell line was cultured in Dulbecco's modified Eagle's medium (Life Technologies) containing 10% fetal bovine serum (FBS). The SK-BR-3 cell line was cultured in RMPI-1640 medium (Life Technologies) containing 10% FBS. The BT-20 cell line was cultured in MEM medium (Life Technologies) containing 10% FBS. The MCF-7 cell line was cultured in MEM medium containing 10% FBS, 1 mM sodium pyruvate, 1× non-essential amino acids solution (Life Technologies) and 10 μg/ml insulin (Sigma).

Shigematsu M., Honda S., Loher P., Telonis A.G., Rigoutsos I, & Kirino Y. (2017). YAMAT-seq: an efficient method for high-throughput sequencing of mature transfer RNAs. Nucleic Acids Research, 45(9), e70.

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In Vitro Expansion and Analysis of NK Cells

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NK cell expansion was performed in vitro by using VarioMACS (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and cultured in MEM medium (Thermo Fisher Scientific, Inc., Waltham, MA, USA) for 72 h at 37°C. On day 5, MEM medium (Thermo Fisher Scientific, Inc.) was refreshed with the addition of autoplasma (1%) (Sigma-Aldrich; Merck KGaA) and interleukin-2 (500 IU/ml) (Sigma-Aldrich; Merck KGaA). Culture continued for 14 days at 37°C. Cells were then fixed with 4% paraformaldehyde for 15 min at 37°C and the viability of expanded NK cells was determined by 5% propidium iodide staining for 2 h at 37°C as described previously (25 (link)). In vitro-expanded NK cells were stained with primary antibodies and analyzed using a flow cytometer with antibodies and FCS Express^™ 4 IVD software (De Novo Software, Glendale, CA, USA), as described in a previous study (24 (link)).

Sun Y., Yao Z., Zhao Z., Xiao H., Xia M., Zhu X., Jiang X, & Sun C. (2018). Natural killer cells inhibit metastasis of ovarian carcinoma cells and show therapeutic effects in a murine model of ovarian cancer. Experimental and Therapeutic Medicine, 16(2), 1071-1078.

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Sourcing and Culturing Cell Lines

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Neonatal human dermal fibroblasts (NDFs) were purchased from ATCC (ATCC, PCS-201-010, Manassas, VA, USA). Human iPSC endothelial cells were purchased from FUJIFILM (FUJIFILM Cellular Dynamics, R1022, WI, USA) and maintained in iCell endothelial cells medium (Supplemental Table 1). Neonatal human epidermal keratinocytes (NHEKs) were purchased from Sciencell (Sciencell,2100, Carlsbad, CA, USA). NDFs were maintained in MEM medium supplemented with 5 % FBS (Thermo Fisher Scientific, Waltham, MA, USA). NHEKs were maintained in keratinocyte growth medium (KGM) purchased from Sciencell (Sciencell, 2101, Carlsbad, CA, USA). Human placental microvascular pericytes were purchased from agio-protemomie(Angio-proteomie, cAP-0029, Boston, MA, USA) and cultured in MEM medium supplemented with 5 % FBS (Thermo Fisher Scientific, Waltham, MA, USA). All the cells were maintained at 37 °C under a 5 % CO₂ atmosphere. Cells of passage number 3-4 were used. All the cultures were routinely monitored for mycoplasma contamination using MycoAlert PLUS mycoplasma detection kit (Lonza, Walkersville, MD, USA).

Liu X., Michael S., Bharti K., Ferrer M, & Song M.J. (2020). A Biofabricated Vascularized Skin Model of Atopic Dermatitis for Preclinical Studies. Biofabrication, 12(3), 035002.

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Cell Culture Conditions for Bladder Cancer Lines

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SV-HUC-1, 5637, J82, TCC-SUP, and UMUC-3 bladder cell lines were purchased from the American Type Culture Collection. SV-HUC-1 cell line was maintained in Ham’s F12K medium (Thermo Scientific) supplemented with 10% fetal bovine serum, 5637 was maintained in RPMI 1640 medium (Thermo Scientific) supplemented with 10% fetal bovine serum and 5% glutamine, J82 was maintained in MEM medium (Thermo Scientific) supplemented with 10% fetal bovine serum and 5% glutamine, and TCC-SUP and UMUC-3 were maintained in MEM medium (Thermo Scientific) supplemented with 10% fetal bovine serum and 5% glutamine. These cell lines were grown at 37°C containing 5% CO₂ in a humidified chamber.

Qiu D., Zhu Y, & Cong Z. (2020). YAP Triggers Bladder Cancer Proliferation by Affecting the MAPK Pathway. Cancer Management and Research, 12, 12205-12214.

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Culturing Breast Cancer Cell Lines

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MCF7, SK-BR3, MDA-MB231 and MDA-MB-468 cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA). MCF7 was cultured in MEM medium (Gibco – Life Technologies, Carlsbad, CA, USA) containing a final concentration of 0.01 mg/ml human recombinant insulin. SK-BR3 was cultured in MCoy 5A medium (Hyclone, UT, USA). MDA-MB-231 and MDA-MB-468 were cultured in MEM medium (Gibco). All media were supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 units/mL penicillin G, and 100 μg/mL streptomycin sulfate and grown in a humidified incubator at 37°C and 5% CO₂ atmosphere. Bafilomycin A1 (Millipore, Darmastabt, Germany), Concanamycin A (Sigma-Aldrich, St. Louis, MO, USA) and Gamma Secretase inhibitor (GSI-IX, Millipore) were dissolved in DMSO and used at the indicated concentrations.

Pamarthy S., Jaiswal M.K., Kulshreshtha A., Katara G.K., Gilman-Sachs A, & Beaman K.D. (2015). The Vacuolar ATPase a2-subunit regulates Notch signaling in triple-negative breast cancer cells. Oncotarget, 6(33), 34206-34220.

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Culturing Diverse Lung Cancer Cell Lines

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The human lung adenocarcinoma cell lines (A549, H157, H1975, H2228, H3122, H2030, and H1299), and human large cell lung cancer cell line H460 were cultured in RPMI-1640 medium (GIBCO); the human squamous cell lung carcinoma cells (SK-MES-1) were cultured in MEM medium (GIBCO); and the human normal embryonic lung fibroblast cell line IMR-90 was cultured in MEM medium supplemented with 10% FBS (GIBCO), GlutaMAX (GIBCO), MEM non-essential amino acids solution (GIBCO), sodium pyruvate (GIBCO), and penicillin–streptomycin solution. All cells were maintained in a humidified chamber at 37 °C in 5% CO₂.

Li C., Qin F., Hu F., Xu H., Sun G., Han G., Wang T, & Guo M. (2018). Characterization and selective incorporation of small non-coding RNAs in non-small cell lung cancer extracellular vesicles. Cell & Bioscience, 8, 2.

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Human Colon Cancer Cell Lines

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Human colon adenocarcinoma cell line SW480, LoVo, HCT-116, Caco2 and SW620 were obtained from the Central Laboratory of Medical College of Xi’an Jiaotong University. Cell lines SW480, LoVo, HCT-116 and SW620 were cultured in RPMI 1640 medium (Gibco, CA, USA), and the Caco2 cell line was cultured in MEM medium (Gibco, CA, USA) with 10% fetal bovine serum (FBS; Gibco, CA, USA) at 37°C in a humidified incubator with 5% CO₂. Next, 0.25% of pancreatin (Hyclone, Utah, USA) with 0.05% ethylenediaminetetraacetic acid (EDTA) was used to detach cells for subculture. Drug resistant human colon carcinoma SW480/5-FU and LoVo/5-FU cell lines were purchased from Keygen Biotech (Nanjing, China), and cultured in MEM medium (Gibco, CA, USA), containing 25 μg/ml of 5-FU (Sigma, St. Louis, USA) and 10% FBS (pH 7.4), under the same conditions as the SW480 cell line.

Chen N., Kong Y., Wu Y., Gao Q., Fu J., Sun X, & Geng Q. (2019). CAC1 knockdown reverses drug resistance through the downregulation of P-gp and MRP-1 expression in colorectal cancer. PLoS ONE, 14(9), e0222035.

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Propagation of DENV Strains in Aedes Cell Line

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DENV strains in our study were isolated from the positive serums of patients, and the Aedes albopictus gut cell line (C6/36) was used for virus propagation. Serum samples were diluted 10-fold with fresh MEM medium (Gibco, Waltham, MA, USA) and then inoculated with a C6/36 cell monolayer, which was incubated in MEM medium supplemented with 2% fetal bovine serum (FBS, Gibco, USA) at 28°C for 5–7 days. Cells showed typical cytopathic effects (CPE) being considered DENV positive. These culture supernatants were then stored at −80°C for use.

Yao W., Yang Z., Lou X., Mao H., Yan H, & Zhang Y. (2021). Molecular Characterization of Dengue Virus Type 1 in Zhejiang in 2019. Frontiers in Cellular and Infection Microbiology, 11, 673299.

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STIM1 Overexpression in HEK 293 Cells

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All cells were cultivated at 37 °C and 5% CO₂. MEM medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FCS (Gibco) was used for HEK 293 WT cells. HEK 293 cells stably expressing STIM1 were grown in MEM medium supplemented with 10% FCS and 500 µg/mL G418 (Gibco). All cells were transfected with 1 µg DNA per 10⁶ cells in a Nucleofector 4D device with Nucleofector Kit SF (Lonza, Basel, Switzerland), according to the manufacturer’s guidelines. DNA constructs 1-1-1-1, 3-1-1-1, 3-1-3-1 and 3-1-3-3 were generated previously [33 (link),65 (link)]. All work was performed under the ethics approval of the Bundesamt für Umwelt, Switzerland to the Institute of Biochemistry and Molecular Medicine (A141370).

Kappel S., Kilch T., Baur R., Lochner M, & Peinelt C. (2020). The Number and Position of Orai3 Units within Heteromeric Store-Operated Ca2+ Channels Alter the Pharmacology of ICRAC. International Journal of Molecular Sciences, 21(7), 2458.

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