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Gibco opti mem media

Manufactured by Thermo Fisher Scientific
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Gibco® Opti-MEM™ Media is a cell culture medium designed to support the growth and maintenance of various cell types. It is a serum-free and protein-free formulation that provides the necessary nutrients and components for cell proliferation and survival.

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Lab products found in correlation

Xcelligence dp instrument, by Roche (2 mentions) 16 well cim plate, by Roche (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Lipofectamine transfection reagent, by Thermo Fisher Scientific (1 mentions) Bovine plasma fibronectin, by Merck Group (1 mentions) Anti β actin antibody sc 47778, by Santa Cruz Biotechnology (1 mentions) Iso hydrochloride, by Merck Group (1 mentions) Anti rabbit and anti mouse secondary antibody, by Cell Signaling Technology (1 mentions) Mitosox red, by Thermo Fisher Scientific (1 mentions) N acetyl cysteine (nac), by Merck Group (1 mentions) Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (1 mentions) Curcumin, by Merck Group (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Invitrogen lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Opti-MEM (8992 citations) Opti-MEM media (656 citations) MEM media (69 citations) Opti-MEMTM (50 citations) Gibco Opti-MEM (22 citations) Gibco MEM (8 citations) Gibco Opti-MEM reduced serum media (3 citations) Gibco opti-MEM Glutamax reduced serum media (3 citations) Gibco Opti-MEM I Reduced Serum Media (2 citations)

9 protocols using gibco opti mem media

1

Real-Time Monitoring of Cell Migration and Invasion

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These assays were performed using the Roche xCELLigence DP instrument as described previously [24 (link)]. Briefly, for the migration assays, a 16-well CIM plate (Roche) was equilibrated with pre-warmed serum free media (Gibco® Opti-MEM™ Media (Thermo-Fisher Scientific, Australia). Upper and lower chambers of the CIM plate were filled with 160 µL of complete serum medium and placed in 37 °C incubator for 1 h to allow the plate to equilibrate. Approximately, 40,000 cells suspended in 130 µL Gibco® Opti-MEM™ Media (Thermo-Fisher Scientific, Australia) were seeded on the top compartment of the pre-equilibrated 16-well CIM plate (Roche). Readings were taken every 15 min for ~ 80 h. Each plate contained two duplicate wells and each experiment was repeated 3 times, the mean results were illustrated graphically using PRISM software.
For invasion assays, the CIM plates were coated with 20 µL of Matrigel. Cells (4 × 104) suspended in 130 µL Gibco® Opti-MEM™ Media (Thermo-Fisher Scientific, NSW, Australia) were seeded on the top compartment of the pre-equilibrated 16-well CIM plate (Roche). Readings were taken every 15 min for ~ 40 h. Each plate contained two duplicate wells and each experiment was repeated 3 times. Linear regression analysis of two slopes arising from Cont and T2-KD cells were used to assess significance.
Escalona R.M., Chu S., Kadife E., Kelly J.K., Kannourakis G., Findlay J.K, & Ahmed N. (2022). Knock down of TIMP-2 by siRNA and CRISPR/Cas9 mediates diverse cellular reprogramming of metastasis and chemosensitivity in ovarian cancer. Cancer Cell International, 22, 422.
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2

HEK293 Cell Culture and Heterologous Protein Expression

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HEK293 cells were cultured in minimum essential medium supplemented with 1% nonessential amino acid solution, 10% horse serum, 1% sodium pyruvate solution, and 1.4% penicillin/streptomycin solution. All cells were plated in T25 flasks and stored in a 5% CO2 incubator at 37°C for 24 hours. Heterologous expression of Kv4.3 and SEMA3A was accomplished by co-transfecting 0.5 μg of pIreGFP-KCND3WT with 1.0 μg pIRES2-dsRed2-SEMA3AWT or pIRES2-dsRed2-SEMA3AR552C or pIRES2-dsRed2-SEMA3AR734W (0.5 μg of WT-SEMA3A + 0.5 μg of mutant-SEMA3A were used for heterozygote co-expression studies) using 5 μL of Lipofectamine transfection reagent (Invitrogen, Carlsbad, CA) in Gibco® OPTI-MEM media (Invitrogen, Carlsbad, CA). Cells fluorescing 48 hours post-transfection were selected for electrophysiological experiments. HEK293 cell culture and transfection procedures for Nav1.5, Cav1.2 and Kv4.2 are included in the online data supplement.
Boczek N.J., Ye D., Johnson E.K., Wang W., Crotti L., Tester D.J., Dagradi F., Mizusawa Y., Torchio M., Alders M., Giudicessi J.R., Wilde A.A., Schwartz P.J., Nerbonne J.M, & Ackerman M.J. (2014). Characterization of SEMA3A-Encoded Semaphorin as a Naturally Occurring Kv4.3 Protein Inhibitor and its Contribution to Brugada Syndrome. Circulation research, 115(4), 460-469.
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3

Glioblastoma Cell Motility on STEP Fibers

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Glioblastoma U251 cells stably expressing GFP-Actin16 (link) were cultured in Gibco® Opti-MEM® media (Invitrogen Corporation, Carlsbad, CA, USA) containing 10% fetal bovine serum. Four hundred nm diameter polystyrene STEP Fiber substrates18 (link) were attached to MatTek No. 0 glass bottom 35 mm dishes (MatTek Corp., Ashland, MA, USA) onto Dow-Corning® High Vacuum Grease (Dow-Corning Inc., Midland, MI, USA). The STEP Fiber substrate was incubated with 1 mg/mL bovine plasma fibronectin (Sigma-Aldrich Inc., St. Louis, MO, USA) for 4 h at 37 °C prior to cell seeding. 50 μL of cell suspension was seeded onto fibronectin-coated STEP fiber substrates at a concentration of 100,000 cells/mL and incubated for 3 h prior to imaging.
Estabridis H.M., Jana A., Nain A, & Odde D.J. (2017). Cell Migration in 1D and 2D Nanofiber Microenvironments. Annals of Biomedical Engineering, 46(3), 392-403.
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4

Cardioprotective Effects of Antioxidants

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H9C2 rat cardiomyoblast cells obtained from the Korean Cell Line Bank (Seoul, Korea) were maintained in a humidified atmosphere at 37 °C and 5% CO2 in Dulbecco’s modified Eagle’s medium containing 1 g/L glucose (L-DMEM; Lonza, Switzerland) supplemented with 10% fetal bovine serum (FBS; Serena, Germany) and 1% penicillin/streptomycin. Cells were differentiated over 7 days by incubation in media containing 1% FBS that was changed daily. Experiments were performed 24 to 48 h after seeding of differentiated cells by adding compounds to culture medium. N-Acetyl-L-cysteine (NAC) and curcumin was added 1 h prior to initiation of ISO treatment.
ISO hydrochloride, NAC, and curcumin were from Sigma Aldrich (St. Louis, MO). MitoSOX Red, Lipofectamine RNAi MAX reagent, and GIBCO Opti-MEM media were from Invitrogen Life Technologies (Carlsbad, CA). Antibodies against phosphorylated extracellular-signal-regulated kinase-1/2 (ERK1/2; #9101S), total ERK2 (#9108S) and p53 (#9282S), and anti-rabbit and anti-mouse secondary antibodies were from Cell Signaling Technology (Danvers, MA). Non-targeted small interfering RNA (siRNA), siRNA against p53, and anti-β-actin antibody (#sc-47778) were from Santa Cruz Biotechnology (Dallas, TX, USA).
Lee J.H., Kim D.H., Kim M., Jung K.H, & Lee K.H. (2022). Mitochondrial ROS-Mediated Metabolic and Cytotoxic Effects of Isoproterenol on Cardiomyocytes Are p53-Dependent and Reversed by Curcumin. Molecules, 27(4), 1346.
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5

Cardiac Fibroblast CTGF miRNA Transfection

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Adult cardiac fibroblast (30,000 cells on 12 well plate) were plated and the following day transiently transfected with Lipofectamine 3000 (Thermo Fisher Scientific, catalog L3000015) and CTGF miRNA (GeneCopoeia) in Gibco Opti-MEM media (Thermo Fisher Scientific). Cells were treated with 50 nM miR-18a-5p mimic or inhibitor as described previously. The luciferase assay was performed using the GeneCopoeia Luc-Pair Duo-Luciferase Assay Kit 2.0 (GeneCopoeia). Cells were prepared and washed according to the manufacturer’s recommendations. The ratio of luminescence was measured on luminometer.
Kraus L., Ma L., Yang Y., Nguyen F., Hoy R.C., Okuno T., Khan M, & Mohsin S. (2020). Cortical Bone Derived Stem Cells Modulate Cardiac Fibroblast Response via miR-18a in the Heart After Injury. Frontiers in Cell and Developmental Biology, 8, 494.
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6

Sab siRNA Transfection Protocol

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The protocol of Sab small interfering RNA is similar to our previous study [21 (link)]. The cells were inoculated in 6-well cell culture dishes and transfected with Sab siRNA by Gibco™ Opti-MEM™ media (Thermo-Fisher Scientific, Waltham, MA, USA). In brief, 3 μl of 20 μM Sab siRNA-1, 3 μl of 20 μM Sab siRNA-2, and 4 μl of 20 μM Sab siRNA-3—that is, Sab siRNA-1 : Sab siRNA-2 : Sab siRNA-3 = 3 : 3 : 4—or 10 μl negative control (NC) siRNA were mixed with 250 μl of the Opti-MEM™ media, separately (the sequences of each siRNA and negative control were shown in Table 1). Next, Lipofectamine™ 2000 (Invitrogen, Carlsbad, CA, USA) was mixed with the Opti-MEM™ media, and then, the mixtures were combined for 20 minutes at room temperature. Subsequently, H9c2 cells were incubated with mixture for 6 hours. At last, the medium was then changed into antibiotic-free DMEM supplemented with 10% FBS for 48 hours.
Chu Q., Zhang Y., Zhong S., Gao F., Chen Y., Wang B., Zhang Z., Cai W., Li W., Zheng F, & Shi G. (2019). N-n-Butyl Haloperidol Iodide Ameliorates Oxidative Stress in Mitochondria Induced by Hypoxia/Reoxygenation through the Mitochondrial c-Jun N-Terminal Kinase/Sab/Src/Reactive Oxygen Species Pathway in H9c2 Cells. Oxidative Medicine and Cellular Longevity, 2019, 7417561.
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7

Investigating miR-18a-5p Modulation in Cardiac Fibroblasts

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Adult cardiac fibroblasts were transiently transfected with 50 nM miR-18a-5p mimic (Thermo Fisher Scientific, catalog 4464066), miR-18a-5p inhibitor (Thermo Fisher Scientific, catalog MH12973), or negative control (Thermo Scientific, catalog 4464058) using Invitrogen Lipofectamine 3000 (Thermo Fisher Scientific, catalog L3000015) in serum free Gibco Opti-MEM media (Thermo Fisher Scientific, catalog 31985062) according to the manufacturer’s recommendations. After 24 h post transfections, cells were used for protein and RNA analysis or stimulated with 10ng TGF-β for 48 h before harvesting.
Kraus L., Ma L., Yang Y., Nguyen F., Hoy R.C., Okuno T., Khan M, & Mohsin S. (2020). Cortical Bone Derived Stem Cells Modulate Cardiac Fibroblast Response via miR-18a in the Heart After Injury. Frontiers in Cell and Developmental Biology, 8, 494.
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8

Cell Invasion Assay Using xCELLigence

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Cell invasion assay was performed by using the Roche xCELLigence DP instrument as described previously [35 ]. The upper chamber of the 16-well CIM plate (Roche, NSW, Australia) was coated with 20 μl of matrigel and left to set for 30 min at 37 °C. Cells (4X104) suspended in 130ul Gibco® Opti-MEM™ Media (Thermo-Fisher Scientific, NSW, Australia) were seeded on the top compartment of the pre-equilibrated 16-well CIM plate (Roche). Readings were taken every 15 min for ~ 40 h. Each plate contained two duplicate wells and each experiment was repeated 3 times. Mean values from three experiments on each Cont and T2-KD cell lines are illustrated graphically using PRISM software. Linear regression analysis of two slopes arising from Cont and T2-KD cells were used to obtain significant values.
Escalona R.M., Bilandzic M., Western P., Kadife E., Kannourakis G., Findlay J.K, & Ahmed N. (2020). TIMP-2 regulates proliferation, invasion and STAT3-mediated cancer stem cell-dependent chemoresistance in ovarian cancer cells. BMC Cancer, 20, 960.
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9

Murine Embryonic Fibroblast Generation

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MEF cells were generated using E12.5-14.5 embryos as described in Takahashi and Yamanaka (2006) (link), using Gibco Opti-MEM media (Thermo Fisher Scientific) with 10% fetal calf serum, and grown in hypoxic conditions (5% CO2, 3% O2). Cells were passaged three times before use, and littermate controls were used where possible.
Findlay A.S., Carter R.N., Starbuck B., McKie L., Nováková K., Budd P.S., Keighren M.A., Marsh J.A., Cross S.H., Simon M.M., Potter P.K., Morton N.M, & Jackson I.J. (2018). Mouse Idh3a mutations cause retinal degeneration and reduced mitochondrial function. Disease Models & Mechanisms, 11(12), dmm036426.
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