Tissue culture plates

Manufactured by Greiner

Sourced in Austria, Germany

Tissue culture plates are a type of laboratory equipment used for the cultivation and growth of cells, tissues, or organisms in a controlled environment. They provide a stable and sterile platform for culturing biological samples.

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Lab products found in correlation

Celltiter 96 aqueous one solution cell proliferation assay mts reagent, by Promega (2 mentions) Spectramax plus 384, by Molecular Devices (2 mentions) Recombinant human tg2, by R&D Systems (1 mentions) Bisphenol c bpc, by Merck Group (1 mentions) Bisphenol af bpaf, by Merck Group (1 mentions) Luciferin, by Promega (1 mentions) Incucyte, by Sartorius (1 mentions) Celltiter 96 aqueous one solution cell proliferation assay, by Promega (1 mentions) Novocyte, by Agilent Technologies (1 mentions) Thioglycollate medium, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Glutamate, by Merck Group (1 mentions) Sh sy5y cell, by American Type Culture Collection (1 mentions) 3 morpholinosydnonimine, by Merck Group (1 mentions) Sin 1, by Merck Group (1 mentions)

11 protocols using tissue culture plates

Preparation and Coating of TG2-cross-linked Fibrinogen

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TG2-cross-linked fibrinogen was prepared as described previously with minor modifications [12 (link)]. Human fibrinogen 1 (Enzyme Research Labs, South Bend, IN, 2.3 μM) was incubated with recombinant human TG2 (R&D Systems, Minneapolis, MN, 0.3 μM) in buffer containing 5 mM CaCl₂, 50 mM Tris-HCl, and 274 mM NaCl at 37 °C for 1 hour. Unmodified fibrinogen was prepared under the same conditions, by incubating in buffer containing 5 mM CaCl₂, 50 mM Tris-HCl, and 274 mM NaCl at 37 °C for 1 hour, in the absence of TG2. In some experiments, the transglutaminase inhibitor cystamine (Cayman Chemical, Ann Arbor, MI, 0–2.5 mM) was added to the reaction. Purified mouse fibrinogen (Enzyme Research Labs) was used in select experiments, where indicated.
To coat tissue culture plates, fibrinogen and TG2-cross-linked fibrinogen were diluted to 10 μg/mL in sterile phosphate-buffered saline (PBS) and 1 mL, 0.5 mL, or 0.2 mL of solution was added to the wells of 6, 12, or 24-well tissue culture plates (Greiner Bio-One, Monroe, NC), respectively. Plates were incubated overnight at 4 °C and rinsed 3 times with PBS before plating cells.

Poole L.G., Kopec A.K., Flick M.J, & Luyendyk J.P. (2022). Cross-linking by tissue transglutaminase-2 alters fibrinogen-directed macrophage proinflammatory activity. Journal of thrombosis and haemostasis : JTH, 20(5), 1182-1192.

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Cytotoxicity Screening of Compounds

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Adherent cell lines Hap-1 and Capan-1 cells were seeded at a density of 500 cells per well, whereas HCT-116 and NCI-H460 were seeded at 1500 cells per well in 384-well tissue culture plates (Greiner, Kremsmünster, Austria). After overnight incubation, cells were treated with seven different concentrations of the test compounds, ranging from 100 to 0.006µM. Suspension cell lines HL-60, K-562, Z-138, and MM.1S were seeded at 2500 cells per well, and for the DND-41 cell line, 5500 cells were seeded in 384-well culture plates containing the test compounds at the same concentration points. Cells were incubated for 72 h with compounds and were then analyzed using the CellTiter 96^® AQueous One Solution Cell Proliferation Assay (MTS) reagent (Promega, Fitchburg, WI, USA) according to the manufacturer’s instructions. The absorbance of the samples was measured at 490 nm using a SpectraMax Plus 384 (Molecular Devices, San Jose, CA, USA), and OD values were used to calculate the 50% inhibitory concentration (IC₅₀). Compounds were tested in at least two independent experiments.

Perin N., Gulin M., Kos M., Persoons L., Daelemans D., Fabijanić I., Stojković M.R, & Hranjec M. (2024). Synthesis and Biological Evaluation of Novel Amino and Amido Substituted Pentacyclic Benzimidazole Derivatives as Antiproliferative Agents. International Journal of Molecular Sciences, 25(4), 2288.

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Endocrine Disruptor Compound Screening

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Tissue culture plates were obtained from Greiner Bio-one (Monroe, NC, USA), and media was purchased from Invitrogen (Grand Island, NY, USA). Luciferin (sodium salt) was purchased from Promega (Charbonnières, France). The chemicals 17β-estradiol (E2), 17α-ethinylestradiol (EE2), bisphenol A (CAS no. 80–05-7), bisphenol AF (BPAF; CAS no. 1478–61-1), and bisphenol C (BPC; CAS no. 14868–03-2) were purchased from Sigma-Aldrich (Saint-Quentin Fallavier, France). ICI 182,780 (7α,17β-[9[(4,4,5,5,5-pentafluoropentyl)sulfinyl]nonyl]estra-1,3,5(10)-triene-3,17-diol, fulvestrant) was obtained from Zeneca (Macclesfield, UK), and 3,17-Dihydroxy-19-nor-17α-pregna-1,3,5(10)-triene-21,16α-lactone (16α-LE2) was a kind gift of Peter Muhn from Research Laboratories of Schering AG (Berlin, Germany). Compounds were dissolved in dimethyl sulfoxide (DMSO) as 10⁻² M stock solutions.

Pinto C., Hao R., Grimaldi M., Thrikawala S., Boulahtouf A., Aït-Aïssa S., Brion F., Gustafsson J.Å., Balaguer P, & Bondesson M. (2019). Differential activity of BPA, BPAF and BPC on zebrafish estrogen receptors in vitro and in vivo. Toxicology and applied pharmacology, 380, 114709.

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Chick DRG Neurite Outgrowth Assay

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Twenty-four-well tissue culture plates (Greiner) were coated with 50 µg/ml poly-L-lysine and 8 µg/ml mouse laminin. DRG were dissected from E7 or E8 chick embryos and placed in a well containing 500 µl of F12 Ham’s medium supplemented with 2% fetal bovine serum, 16.7 ng/ml 2.5 S NGF, 50 U/ml penicillin, and 50 µg/ml streptomycin. Explants were infected with recombinant herpes-simplex viruses harboring the EGFP-CRMP2-wild type or mutants. Negative controls were infected with HSV expressing EGFP. Explants were cultured at 37 °C for one day. More than 50% of neurites showed EGFP-fluorescence. Neurites were immunostained with anti-α-tubulin rat monoclonal antibodies and Alexa594-conjugated anti-rat IgG. Entire images of neurites from each DRG explant were captured through ×2 objective lens equipped with an Olympus IX-70 microscope and a Spot-2e CCD (Spot) camera.

Niwa S., Nakamura F., Tomabechi Y., Aoki M., Shigematsu H., Matsumoto T., Yamagata A., Fukai S., Hirokawa N., Goshima Y., Shirouzu M, & Nitta R. (2017). Structural basis for CRMP2-induced axonal microtubule formation. Scientific Reports, 7, 10681.

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High-Throughput Cell Viability Assay

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Adherent cell lines LN-229, Capan-1, HCT-116 and NCI-H460 cells were seeded at a density between 500 and 1500 cells per well, in 384-well tissue culture plates (Greiner). After overnight incubation, cells were treated with the test compounds. After incubation for 3 days at 37 °C, the formazan-based 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) cell viability assay (CellTiter 96^® AQueous One Solution Cell Proliferation Assay, Promega) was performed to assess cell viability, and the spectrophotometric data were used to calculate the IC₅₀ values [61 (link)–65 (link)].
Suspension cell lines DND-41, HL-60, K-562 and Z-138 were seeded at densities ranging from 2500 to 5500 cells per well in 384-well tissue culture plates containing the test compounds at the same concentration points. The plates were incubated at 37 °C and monitored for 72 h in an IncuCyte^® device (Essen BioScience Inc.) for real-time imaging. Images were taken every 3 h, with one field imaged per well under 10 × magnification. Cell growth was then quantified based on the percent cellular confluence as analyzed by the IncuCyte^® image analysis software and used to calculate IC₅₀ values by linear interpolation.

Beus M., Persoons L., Daelemans D., Schols D., Savijoki K., Varmanen P., Yli-Kauhaluoma J., Pavić K, & Zorc B. (2022). Anthranilamides with quinoline and β-carboline scaffolds: design, synthesis, and biological activity. Molecular Diversity, 26(5), 2595-2612.

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3T3-L1 Adipocyte Differentiation Assay

Cited 1 time

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We performed the
differentiation assays with 3T3-L1 cells in accordance with a previously
described method.^{28 (link)} In brief, an experiment
consists of 3 days predifferentiation (1 day seeding and 2 days to
allow cells to enter the resting state), followed by an 8 days differentiation
window (2 days induction of differentiation and 6 days maintenance).
Subconfluent cells of passage 10 were trypsinized and counted with
a flow cytometer (NovoCyte, Acea Biosciences). 15,000 cells well^–1 were seeded in 200 μL of preadipocyte medium
(PAM: DMEM-high supplemented with 10% bovine calf serum and 1% penicillin/streptomycin)
into 96-well black, clear-bottom tissue culture plates (655,090, Greiner
Bio-One) and incubated at 37 °C and 5% CO₂. After
24 h, we checked that the cells had reached confluency, replaced the
medium with 200 μL fresh PAM well^–1, and cultured
the cells for another 48 h to initiate growth arrest. We included
a preadipocyte control (undifferentiated cells) on every plate which
we kept cultivated in PAM, while the rest of the cells were differentiated
as described below.

Völker J., Ashcroft F., Vedøy Å., Zimmermann L, & Wagner M. (2022). Adipogenic Activity of Chemicals Used in Plastic Consumer Products. Environmental Science & Technology, 56(4), 2487-2496.

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Cell-Based Cytotoxicity Assay Protocol

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Adherent cell lines LN-229, HCT-116, and NCI-H460 and Capan-1 cells were seeded at a density between 500 and 1500 cells per well, in 384-well tissue culture plates (Greiner, Kremsmünster, Austria). After overnight incubation, the cells were treated with seven different concentrations of the test compounds, ranging from 100 to 0.006 µM.
Suspension cell lines HL-60, K-562, Z-138, and DND-41 were seeded at densities ranging from 2500 to 5500 cells per well in 384-well culture plates containing the test compounds at the same concentration points. The cells were incubated for 72 h with the compounds and were then analyzed using the CellTiter 96^® AQueous One Solution Cell Proliferation Assay (MTS) reagent (Promega, Madison, WI, USA) according to the manufacturer’s instructions. The absorbance of the samples was measured at 490 nm using a SpectraMax Plus 384 (Molecular Devices, Silicon Valley, CA, USA), and OD values were used to calculate the 50% inhibitory concentration (IC₅₀). The compounds were tested in two independent experiments.

Beč A., Cindrić M., Persoons L., Banjanac M., Radovanović V., Daelemans D, & Hranjec M. (2023). Novel Biologically Active N-Substituted Benzimidazole Derived Schiff Bases: Design, Synthesis, and Biological Evaluation. Molecules, 28(9), 3720.

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Culturing Lipid-Loaded Foam Cells

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Five mice per group from the atherosclerosis experiment were injected with thioglycollate medium (3%, Fischer). Four days after injection, mice were sacrificed and the peritoneum was flushed once with 10 ml ice-cold PBS to collect peritoneal macrophages (PEMs). Since these mice are on a HFD, these elicited macrophages are fully loaded with lipids and can thus be considered foam cells (21 (link)). Flushed thioglycollate-elicited foam cells were pooled per group and cultured at a density of 5 × 10⁵ cells/well in 24-well tissue culture plates in triplicate (Greiner Bio-One) and cultured in RPMI-1640 with 25 mM HEPES, 2 mM L-glutamine, 10% FCS, penicillin (100 U/ml), and streptomycin (100 μg/ml) (all Gibco) and allowed adherence for 3 h. Hereafter the cells were washed and stimulated for 24 h with LPS (10 ng/ml), LPS plus IFNγ (100 U/ml), IL-4 (20 ng/ml), or left unstimulated.

Neele A.E., Chen H.J., Gijbels M.J., van der Velden S., Hoeksema M.A., Boshuizen M.C., Van den Bossche J., Tool A.T., Matlung H.L., van den Berg T.K., Lutgens E, & de Winther M.P. (2021). Myeloid Ezh2 Deficiency Limits Atherosclerosis Development. Frontiers in Immunology, 11, 594603.

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Motor Neuron-Neuroblastoma Fusion Cell Line Protocol

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Mouse NSC-34 hybrid cell line, a widely used motor neuron-neuroblastoma fusion line was chosen, as it expressed many of the morphological and physiological properties of motor neurons [26 (link)]. Cellswere grown in Dulbecco’s modified Eagle’s (DMEM)/F-12 medium supplemented with 10% fetal calf serum and 1% of a mixture of penicillin/streptomycin at 37°C in an atmosphere of 5% CO2 in air. Human neuroblastoma cells, SH-SY5Y cells were obtained from the ATCC (Rockville, USA), and were grown on tissue culture plates (Greiner, Hessle, UK) in DMEM, supplemented with 10% fetal calf serum, 1% L-glutamine and 1% SPN antibiotics (Biological Industries, Bet Haemek, Israel). Cells were incubated at 37°C in a humidified atmosphere with 5% CO₂. Cells were exposed to peroxynitrite ion generator 3-morpholinosydnonimine (SIN-1, Sigma–Aldrich) or to glutamate (Sigma—Aldrich).

Lev N., Barhum Y., Lotan I., Steiner I, & Offen D. (2015). DJ-1 Knockout Augments Disease Severity and Shortens Survival in a Mouse Model of ALS. PLoS ONE, 10(3), e0117190.

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Tissue Culture Protocol with Reagents

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RPMI 1640 tissue culture medium, penicillin and streptomycin were purchased from Biochrom (Berlin, Germany). Tissue culture plates were obtained from Greiner Bio-One (Frickenhausen, Germany). Custom designed PCR primers and Taqman probes were synthesized from Life Technologies (Karlsruhe, Germany). Palmitoleic acid was obtained from Larodan (Malmö, Sweden). All other chemicals were purchased from Sigma-Aldrich (Taufkirchen, Germany).

Gehrmann W., Würdemann W., Plötz T., Jörns A., Lenzen S, & Elsner M. (2015). Antagonism Between Saturated and Unsaturated Fatty Acids in ROS Mediated Lipotoxicity in Rat Insulin-Producing Cells. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 36(3).

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