Cells were cultured under standard cell culture conditions, 37 °C, 5% CO2, humidified atmosphere. A2780 cells were cultured in RMPI 1640 medium supplemented with 10% fetal calf serum, 2 mM glutamine, and 1% penicillin–streptomycin (Sigma-Aldrich). ID8 cells were cultured in high-glucose DMEM (4.5 g/L glucose) supplemented with 4% fetal calf serum, 2 mM glutamine, 1% penicillin–streptomycin (Sigma-Aldrich), and 1% ITS supplement (Sigma-Aldrich I3146). U251 cells were maintained in MEM (Sigma-Aldrich), 10% fetal bovine serum (Sigma-Aldrich), 1% penicillin/streptomycin (Invitrogene), and 2 mM glutamine. MCF7 cells were maintained in MEM (Sigma-Aldrich), 10% fetal bovine serum (Sigma-Aldrich), 1% penicillin/streptomycin (Invitrogen, Waltham, MA, USA), and 2 mM glutamine. Capan2 cells were maintained in MEM (Sigma-Aldrich), 10% fetal bovine serum (Sigma-Aldrich), 1% penicillin/streptomycin (Invitrogen), and 2 mM glutamine. Human primary dermal fibroblasts were cultured in low-glucose DMEM (1 g/L glucose) supplemented with 20% fetal calf serum, 2 mM glutamine, and 1% penicillin–streptomycin (Sigma-Aldrich).
Minimum essential medium (mem)
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Japan, Italy, France, Macao, Israel, Sao Tome and Principe, Austria, Canada, India, Argentina
MEM is a laboratory equipment product manufactured by Merck Group. It is a cell culture medium that provides essential nutrients and growth factors to support the cultivation of various cell types in vitro. The core function of MEM is to maintain and promote the growth of cells in a controlled and optimal environment.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (116 mentions)
Fetal bovine serum (fbs), by Merck Group (81 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (67 mentions)
Penicillin, by Thermo Fisher Scientific (44 mentions)
Streptomycin, by Thermo Fisher Scientific (44 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (44 mentions)
Penicillin, by Merck Group (42 mentions)
Streptomycin, by Merck Group (41 mentions)
L glutamine, by Merck Group (40 mentions)
Penicillin streptomycin, by Merck Group (34 mentions)
L glutamine, by Thermo Fisher Scientific (32 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (32 mentions)
Glutamax, by Thermo Fisher Scientific (27 mentions)
Rpmi 1640, by Merck Group (23 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (18 mentions)
Non essential amino acids (neaa), by Merck Group (18 mentions)
Fetal calf serum (fcs), by Merck Group (15 mentions)
Sodium pyruvate, by Merck Group (14 mentions)
Rpmi 1640 medium, by Merck Group (13 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (12 mentions)
598 protocols using minimum essential medium (mem)
1
Cell Culture Conditions Across Cell Lines
2
Viral Infection Protocols for Cell Lines
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The human lung adenocarcinoma cells (A549), the human embryonic kidney cells (HEK293) and the African green monkey epithelial cells (Vero) were grown in Dulbecco's modified Eagle's medium (Sigma‐Aldrich). The Mardin‐Darby canine kidney cells (MDCKII) were grown in minimum essential medium (Sigma‐Aldrich). Dulbecco's modified Eagle's medium and minimum essential medium were supplemented with 10% fetal calf serum (Merck‐Millipore). Cells were cultivated at 37°C and 5% CO2 under constantly humidified conditions.
Human influenza virus A/Puerto Rico/8/34 (PR8; H1N1) wt virus and mutants used in this study were generated by the pHW2000‐based eight plasmid reverse genetics system as described elsewhere (Hoffmann et al.,2000 ). Infection of either A549 or MDCKII cells was performed at a defined multiplicity of infection (MOI) for the indicated times as described previously (Dudek et al., 2010 ).
Human influenza virus A/Puerto Rico/8/34 (PR8; H1N1) wt virus and mutants used in this study were generated by the pHW2000‐based eight plasmid reverse genetics system as described elsewhere (Hoffmann et al.,
3
Melatonin and Luzindole Effects on Mouse Embryo Development
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Female mice were superovulated by an intra-peritoneal (IP) injection of 7.5 IU pregnant mare serum gonadotropin (PMSG, G 4877, Sigma-Aldrich, USA), followed by injection of 7.5 IU human chorionic gonadotropin (HCG, Karma, Germany) 48 hr later (10 (link)).
To ensure the reliability of gestation time, female mice were paired overnight with males of proven fertility (ratio = 1:1) and females with a vaginal plug were selected for the study.
At 42-48 hr after the HCG injection, the embryos were flushed into Ham’s F10 (Sigma, USA) medium under a stereomicroscope (Nikon SMZ-2T, Japan) (11 (link)).
After three washes, embryos with normal develop-mental morphologies were randomly divided into 3 groups including:
To ensure the reliability of gestation time, female mice were paired overnight with males of proven fertility (ratio = 1:1) and females with a vaginal plug were selected for the study.
At 42-48 hr after the HCG injection, the embryos were flushed into Ham’s F10 (Sigma, USA) medium under a stereomicroscope (Nikon SMZ-2T, Japan) (11 (link)).
After three washes, embryos with normal develop-mental morphologies were randomly divided into 3 groups including:
α-MEM medium (Sigma, USA) containing 10% fetal bovine serum (FBS) and 10-9 M melatonin as the melatonin-treated group.
α-MEM medium with serum and without melatonin as the simple media group.
α-MEM medium containing serum and 10-9 M luzindole as the Luzindole-treated group.
4
Culturing Human Lung Cell Lines
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As a growth medium, the Minimum Essential Medium Eagle (MEM, Sigma-Aldrich) commercial solution was enriched with 1 mM of penicillin, streptomycin, and 0.25 mM L-glutamine (Sigma-Aldrich). Two MEM-enriched solutions were used: one with 10% Fetal Bovine Serum (FBS-MEM) and one without (MEM, Sigma-Aldrich). Human epithelial lung cancer cell line (A549, CCL-185, passage number: 1) and fibroblast lung cell line (MRC-5, CCL-171, passage number: 30) were purchased from the American Type Culture Collection (ATCC). In FBS-MEM, cells were maintained as a monolayer culture at 37 °C in a humidified 5% CO2 atmosphere. Cells were sub-cultured according to the protocols recommended to the cell’s phenotype and described previously in Jakubczak et al. [23 (link)]. Cells were detached from the culture flasks using 2 mL of 0.05% trypsin-EDTA solution (trypLE Express, Gibco) for 5 min, suspended in 6 mL of FBS-MEM, and centrifuged for 5 min at 1500 rpm (Universal 32 Tabletop Centrifuge, Hettich, Tuttlingen, Germany). The supernatant was removed, and cell pellets were resuspended in the proper amount of FBS-MEM to obtain the cell density of 1 × 105 cells/mL.
5
DEV Virus Plaque Titration in CEF Cells
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CEF cells were washed once with phosphate-buffered saline (PBS) before being infection with the DEV virus. The inoculum was removed at 2 h post-infection and replenished with either fresh medium or 2% Minimum Essential Medium (MEM)-agarose overlay. The MEM-agarose overlay medium contains MEM (Sigma, St Louis, MO, USA), 2% agarose (Thermo Fisher Scientific, Waltham, MA, USA), 100 units/mL penicillin, 100 µg/mL streptomycin, 2 mM l -glutamine (Sigma), 0.3% bovine serum albumin (BSA) (Sigma), 15 mM 4-(2-hydroxyethyl)-1 -piperazineethanesulfonic acid (HEPES) (Sigma), 0.22% sodium bicarbonate (Sigma) and 0.01% Diethylaminoethyl (DEAE)-Dextran (Sigma). For the virus plaque titration, infected cells were incubated at 37 °C under a 5% CO2 atmosphere for 5 days before being fixed with 1% crystal violet (Sigma) in 20% ethanol for plaque counting and plaque size measurement.
6
Investigating MEM-induced cell proliferation
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MEM (Sigma, MO, USA) was dissolved in 0.9% saline. Mice were injected intraperitoneally with MEM at a dose of 50 mg/kg body weight once a week for 4 weeks. Control mice were injected with the same volume of 0.9% saline. At 2 days after each MEM injection, the mice received a single injection with 50 mg/kg body weight of BrdU (Sigma).
7
Regulation of Endometrial Cancer Cell Lines
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Cells of the EC cell line Ishikawa were cultured in an incubator (5% CO2 at 37°C) with culture medium (MEM [Sigma‐Aldrich], 1% penicillin–streptomycin, 1% non‐essential amino acid [NEAA], 2mM glutamine, and 10% FBS [Gibco, Thermo Fisher Scientific]). The EC cell lines AN3CA and HEC‐1‐B were cultured in an incubator (5% CO2 at 37°C) with culture medium (MEM [Sigma‐Aldrich], 1% penicillin–streptomycin, 1% NEAA, and 10% FBS [Gibco, Thermo Fisher Scientific]). Small interfering RNAs of GAS5, YAP1, PTEN, and miR‐21 mimics/inhibitor were purchased from GenePharma; GAS5 shRNA and overexpression lentivirus were purchased from Genechem. The transfection and transduction were carried out according to the manufacturer’s instructions.
8
Assessing Neurogenesis and Memory in Mice
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MEM (Sigma-Aldrich, St. Louis, MO) was dissolved in 0.9% saline. The mice were injected intraperitoneally (i.p) with MEM at a dose of 25 or 50 mg/kg body weight once a week for four weeks, unless otherwise indicated. The control mice were injected with the same volume of 0.9% saline (vehicle, VEH). As shown in Figure 1B , two days after MEM injection, the mice received single injections with 50 mg/kg body weight of 5-bromo-2-deoxyuridine (BrdU; Sigma-Aldrich). The protein synthesis inhibitor anisomycin (ANI; Wako, Osaka, Japan) was dissolved in artificial cerebrospinal fluid (ACSF), and adjusted to pH 7.4 with NaOH. The sodium channel blocker lidocaine (LIDO, 4%; Sigma-Aldrich) was dissolved in phosphate-buffered saline (PBS) (Frankland et al., 2004 (link)).
9
Culturing Primary Chicken Embryonic Fibroblasts
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Primary chicken embryonic fibroblasts (CEF) were prepared from 10- to 11-day-old chicken embryos (SPF eggs, VALO, Cuxhaven, Germany) and were maintained in Minimum Essential Medium Eagle (MEM) (Sigma-Aldrich, Taufkirchen, Germany) containing 10% heat-inactivated fetal bovine serum (FBS) (Sigma-Aldrich, Taufkirchen, Germany) and 1% MEM non-essential amino acid solution (Sigma-Aldrich, Taufkirchen, Germany). Madin-Darby Canine kidney (MDCK) cells (ATCC CCL-34) were cultured in MEM (Sigma-Aldrich, Taufkirchen, Germany) containing 10% heat-inactivated FBS and 1% Penicillin–Streptomycin (Pen/Strep) (Sigma-Aldrich, Taufkirchen, Germany). Cells were maintained at 37 °C and 5% CO2 atmosphere.
10
Quantifying Salmonella Invasion of Chicken Fibroblasts
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Primary chicken embryo fibroblasts (CEFs) were obtained from 12-dayold specific pathogen free (SPF) embryos of the Leghorn breed. CEFs were cultured in MEM (Sigma-Aldrich) complemented with 5% fetal calf serum (FCS). One day prior to infection, fibroblasts were seeded into 36-mm Petri dishes (Nunc) and grown overnight at 37 °C under 5% CO 2 .
The invasiveness of the Salmonella strains was tested as described by Barrow and Lovell (1989) . Briefly, semi-confluent cell cultures were washed three times with PBS and cultivated in DMEM (Sigma-Aldrich) supplemented with 5% FCS and 1% D-mannose. Overnight bacterial cultures were incubated with fibroblasts at 1:200 dilutions for 2 h at 37 °C under 5% CO 2 . The infectious dose was 4-6 × 10 6 CFU/ml. The number of bacteria in the supernatant was determined by plating serial dilutions on Bromothymol Blue (BTB) agar plates. CEFs were washed three times with PBS and incubated for 1.5 h at 37 °C and 5% CO 2 in MEM (Sigma-Aldrich) containing kanamycin 250 μg/ml to eliminate extracellular bacteria. Finally, cells were washed three times with PBS and digested with 0.025% trypsin (Sigma-Aldrich) and 1% Tween 20 (Sigma-Aldrich) in 0.01M NaH 2 PO 4 (pH 8) for 30 min at 37 °C. Intracellular Salmonella counts were determined by plating serial dilutions on BTB plates. Salmonella invasion was tested three times, with 2-3 replicates each time.
The invasiveness of the Salmonella strains was tested as described by Barrow and Lovell (1989) . Briefly, semi-confluent cell cultures were washed three times with PBS and cultivated in DMEM (Sigma-Aldrich) supplemented with 5% FCS and 1% D-mannose. Overnight bacterial cultures were incubated with fibroblasts at 1:200 dilutions for 2 h at 37 °C under 5% CO 2 . The infectious dose was 4-6 × 10 6 CFU/ml. The number of bacteria in the supernatant was determined by plating serial dilutions on Bromothymol Blue (BTB) agar plates. CEFs were washed three times with PBS and incubated for 1.5 h at 37 °C and 5% CO 2 in MEM (Sigma-Aldrich) containing kanamycin 250 μg/ml to eliminate extracellular bacteria. Finally, cells were washed three times with PBS and digested with 0.025% trypsin (Sigma-Aldrich) and 1% Tween 20 (Sigma-Aldrich) in 0.01M NaH 2 PO 4 (pH 8) for 30 min at 37 °C. Intracellular Salmonella counts were determined by plating serial dilutions on BTB plates. Salmonella invasion was tested three times, with 2-3 replicates each time.
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