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Cell culture plates

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Cell culture plates are a type of laboratory equipment used for the growth and maintenance of cells in controlled environments. They provide a flat, sterile surface for culturing cells and supporting their growth, division, and function.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions) Glutamax, by Thermo Fisher Scientific (4 mentions) Penicillin, by Thermo Fisher Scientific (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) Hek293t, by Thermo Fisher Scientific (2 mentions) Dneasy blood and tissue kit, by Qiagen (2 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions) Salmonella typhosa lps, by Merck Group (1 mentions) Interleukin 4 (il 4), by Merck Group (1 mentions) Trypsin edta 0.5, by Thermo Fisher Scientific (1 mentions) 2 6 dichloro 1 4 benzoquinone, by Merck Group (1 mentions) Cell culture multi well plates, by Thermo Fisher Scientific (1 mentions) 2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions) Crystal violet, by Merck Group (1 mentions) Deuterated water d2o, by Deutero (1 mentions) Geltrex coated, by Thermo Fisher Scientific (1 mentions) Pbs edta, by Thermo Fisher Scientific (1 mentions) E8 supplement, by Thermo Fisher Scientific (1 mentions) Essential 8 medium, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Cellstar® PS 96-well cell culture plates Cell Culture Multi-well Plates 12-well suspension cell culture plates Six-well cell culture plates Black 96-well cell culture plates 384-well white clear-bottom cell culture plates Cell culture dishes and plates F-bottom clear cell culture treated 96-wells plates Cell culture flasks and plates 6-well cell culture plates

17 protocols using cell culture plates

1

CRISPR Base Editor Optimization in HEK293T Cells

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HEK293T (ATCC CRL-3216) cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) plus GlutaMax (Thermo Fisher), supplemented with 10% (v/v) fetal bovine serum (FBS) and 1× penicillin-streptomycin (Thermo Fisher Scientific) at 37°C and 5% CO2. Cells were maintained at confluency below 90% and seeded on 48-well cell culture plates (Greiner). 12-16h after seeding, at approximately 70% confluency, cells were transfected using 1.5μl of Lipofectamine 2000 (Thermo Fisher Scientific) and 500 ng base editor mRNA and 50 ng sgRNA. Cells were incubated for 3 days and genomic DNA was isolated using the DNeasy Blood and Tissue kit (Qiagen) according to the manufacturer’s protocol. For FACS experiments, SaKKH-CBE3 (Addgene #85170) was co-transfected with a control plasmid expressing GFP in HEK293T at 70-80% confluency in 6-well plates (1.5ug BE plasmid, 0.5ug GFP-expressing plasmid, 0.5ug sgRNA-expressing plasmid). An intein-split BE was co-transfected in HEK293T at 70-80% confluency in 6-well plates (N-terminal and C-terminal constructs in a 1:1 ratio, at 1.25ug plasmid each) or the C-terminal part only as RFP control (1.25ug plasmid)5 (link). Different amounts of SaKKH-CBE3 mRNA (4000ng, 200ng, 10ng), together with 400ng mCherry mRNA and 2000ng sgRNA were transfected into HEK293T and Hepa 1-6 cells in a 6-well format and harvested after 48 hours for CBE titration experiments.
Villiger L., Rothgangl T., Witzigmann D., Oka R., Lin P.J., Qi W., Janjuha S., Berk C., Ringnalda F., Beattie M.B., Stoffel M., Thöny B., Hall J., Rehrauer H., van Boxtel R., Tam Y.K, & Schwank G. (2021). In vivo cytidine base editing of hepatocytes without detectable off-target mutations in RNA and DNA. Nature biomedical engineering, 5(2), 179-189.
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2

CRISPR Base Editor Optimization in HEK293T Cells

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HEK293T (ATCC CRL-3216) cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) plus GlutaMax (Thermo Fisher), supplemented with 10% (v/v) fetal bovine serum (FBS) and 1× penicillin-streptomycin (Thermo Fisher Scientific) at 37°C and 5% CO2. Cells were maintained at confluency below 90% and seeded on 48-well cell culture plates (Greiner). 12-16h after seeding, at approximately 70% confluency, cells were transfected using 1.5μl of Lipofectamine 2000 (Thermo Fisher Scientific) and 500 ng base editor mRNA and 50 ng sgRNA. Cells were incubated for 3 days and genomic DNA was isolated using the DNeasy Blood and Tissue kit (Qiagen) according to the manufacturer’s protocol. For FACS experiments, SaKKH-CBE3 (Addgene #85170) was co-transfected with a control plasmid expressing GFP in HEK293T at 70-80% confluency in 6-well plates (1.5ug BE plasmid, 0.5ug GFP-expressing plasmid, 0.5ug sgRNA-expressing plasmid). An intein-split BE was co-transfected in HEK293T at 70-80% confluency in 6-well plates (N-terminal and C-terminal constructs in a 1:1 ratio, at 1.25ug plasmid each) or the C-terminal part only as RFP control (1.25ug plasmid)5 (link). Different amounts of SaKKH-CBE3 mRNA (4000ng, 200ng, 10ng), together with 400ng mCherry mRNA and 2000ng sgRNA were transfected into HEK293T and Hepa 1-6 cells in a 6-well format and harvested after 48 hours for CBE titration experiments.
Villiger L., Rothgangl T., Witzigmann D., Oka R., Lin P.J., Qi W., Janjuha S., Berk C., Ringnalda F., Beattie M.B., Stoffel M., Thöny B., Hall J., Rehrauer H., van Boxtel R., Tam Y.K, & Schwank G. (2021). In vivo cytidine base editing of hepatocytes without detectable off-target mutations in RNA and DNA. Nature biomedical engineering, 5(2), 179-189.
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3

Chemical and Cell Culture Supplies

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Chemicals were purchased from Sigma-Aldrich unless stated otherwise. Cell culture plates were purchased from Greiner Bio-One.
Schirris T.J., Jansen J., Mihajlovic M., van den Heuvel L.P., Masereeuw R, & Russel F.G. (2017). Mild intracellular acidification by dexamethasone attenuates mitochondrial dysfunction in a human inflammatory proximal tubule epithelial cell model. Scientific Reports, 7, 10623.
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4

Fibroblast and Endothelial Cell Culture

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Cell culture plates (24 wells, area 1.9 cm2, Greiner Bio-One, Monroe, NC) were coated with fibronectin (25 μg/ml in phosphate-buffered saline, PBS) for 2 h before cell seeding. Fibroblasts and ECs were cultured separately in serum-free StemSpan, StemPro, and HPGM. M199 containing 1 vol% PSG (penicillin, streptomycin, and L-glutamine) was used as the “standard media” [33 (link), 34 (link), 37 (link)]. ECs were cultured in media with and without the addition of FBS. Fibroblast and ECs were cultured at 35,000 cell/cm2 and 12,000 cell/cm2 density, respectively, at 37°C, 5% CO2 (defined here as “standard conditions”) with media changes on every other day until confluent and ready for testing.
Derakhshan T., Bhowmick R., Ritchey J.W, & Gappa-Fahlenkamp H. (2018). Development of Human Mast Cells from Hematopoietic Stem Cells within a 3D Collagen Matrix: Effect of Stem Cell Media on Mast Cell Generation. Stem Cells International, 2018, 2136193.
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5

Macrophage Differentiation from Murine Bone Marrow

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Bone Marrow was isolated from ORP8 KO male mice (age 10 weeks). Bone marrow cells were plated in DMEM/20% FCS/1% penicillin/1% streptomycin and differentiated into macrophages by addition of 30% L929 cell-conditioned media (as a source of M-CSF) for 7 days according to Van Eck et al. [15] (link) Macrophages were incubated for 24 h in cell culture plates (Greiner Bio-One) the absence or presence of 100 ng/mL LPS (Salmonella Typhosa, Sigma) or 20 ng/mL IL-4 (Sigma) and subsequently lysed for mRNA extraction or analyzed by FACS.
van Kampen E., Beaslas O., Hildebrand R.B., Lammers B., Van Berkel T.J., Olkkonen V.M, & Van Eck M. (2014). Orp8 Deficiency in Bone Marrow-Derived Cells Reduces Atherosclerotic Lesion Progression in LDL Receptor Knockout Mice. PLoS ONE, 9(10), e109024.
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6

Antigen-specific Lymphocyte Proliferation in Pigs

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Antigen-specific lymphocyte proliferation in response to M. hyosynoviae challenge infection was investigated in blood samples from all pigs two days before inoculation and on PID 7 and 12. Proliferation was measured by flow cytometry, assessing cells that had incorporated the thymidine analog Bromo-deoxy-Uridine (BrdU) in newly synthesized DNA (Riber and Jungersen, 2007) . Briefly, peripheral blood mononuclear cells (PBMCs, 3 x 10 6 /ml) in cell culture medium (RPMI 1640 with GlutaMAX™ I, foetal calf serum (10%), penicillin (100 U/ml), streptomycin (100 g/ml)) were incubated in 24 well, cell culture plates (Greiner Labortechnik GmbH, Germany): SEB-culture (Staphylococcal enterotoxin B, 5 g/ml, Alexis, Grünberg, Germany), Ag-culture (Mhyos-antigen, 10 g/ml), RPMI-culture (nil-stimulation).
Incubation was performed for 5 days at 37C in 5% CO 2, the last 18 hours with addition of 5-Brom-2'-Deoxyuridine (BrdU 60 M, Sigma-Aldrich, St. Louis, MO, USA).
Lauritsen K.T., Hagedorn-Olsen T., Jungersen G., Riber U., Stryhn H., Friis N.F., Lind P, & Kristensen B. (2017). Transfer of maternal immunity to piglets is involved in early protection against Mycoplasma hyosynoviae infection. Veterinary immunology and immunopathology, 183.
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7

Synthesis and Characterization of Uremic Toxins

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Chemicals were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands) unless stated otherwise. The uremic toxins p-cresylsulfate and p-cresylglucuronide were synthesized by the Institute for Molecules and Materials, Radboud University, Nijmegen, The Netherlands. MicroPES type TF10 hollow fiber capillary membranes (wall thickness 100 μm, inner diameter 300 μm, max pore size 0.5 μm) were obtained from Membrana GmbH (Wuppertal, Germany). Cell culture plates were purchased from Greiner Bio-One (Monroe, NC).
Jansen J., Fedecostante M., Wilmer M.J., Peters J.G., Kreuser U.M., van den Broek P.H., Mensink R.A., Boltje T.J., Stamatialis D., Wetzels J.F., van den Heuvel L.P., Hoenderop J.G, & Masereeuw R. (2016). Bioengineered kidney tubules efficiently excrete uremic toxins. Scientific Reports, 6, 26715.
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8

DPPH Antioxidant Assay in HaCaT Cells

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All chemicals used were of analytical grade. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) was purchased from Aldrich (Steinheim, Germany). 2,6-dichloro-1,4-benzoquinone was purchased from Sigma Aldrich. HaCaT cells were used for the experiments. Dulbecco′s Modified Eagle Medium (DMEM), Fetal Bovine Serum (FBS), L-Glutamine, penicillin, and streptomycin were purchased from Gibco, Thermo Fischer Scientific (Australia). Trypsin-EDTA 0.5% was purchased from Gibco, Thermo Fischer Scientific (America), and crystal violet was purchased from Merck (Darmstadt, Germany). Cell culture plates were purchased from Greiner Bio-One GmbH (Frickenhausen, Germany). Cell culture multiwell plates were purchased from Thermo Scientific (Denmark). Deuterated water (D2O) and 3-(trimethylsilyl)-1-propionic-2, 2, 3, 3-d4 acid sodium salt (TSP) were obtained from Deutero GmbH (Kastellaun, Germany). Double distilled water (DDW) was used to prepare all solutions. Phosphate Buffer Saline (PBS) was prepared with NaCl: 137.0 mM, KCl: 2.7 mM, Na2HPO4: 10.0 mM, KH2PO4: 1.8 mM. All solutions for the cell cultures were sterilized by autoclaving at 121oC for 20 min, at 21 psi.
Meintani D.G., Chatzimitakos T.G., Kasouni A.I, & Stalikas C.D. (2022). Untargeted metabolomics of human keratinocytes reveals the impact of exposure to 2,6-dichloro-1,4-benzoquinone and 2,6-dichloro-3-hydroxy-1,4-benzoquinone as emerging disinfection by-products. Metabolomics, 18(11), 89.
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9

iPSC Maintenance and Passaging Protocol

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iPSCs (lines 15-0001 and 19) were cultured [as previously described (18 (link))] on Geltrex-coated (ThermoFisher) cell culture plates (Greiner) using Essential™ 8 (E8) medium (Gibco, ThermoFisher) supplemented with E8 supplement (50X, Gibco) and 0.5% (v/v) antibiotic-antimycotic (Gibco). Medium was refreshed daily. Upon 80% confluency, iPSCs were washed using PBS and subsequently split into colonies using PBS-EDTA (0.5 mM, ThermoFisher) for 4 to 6 min at room temperature. Cells were split in a ratio of 1:6—1:8 twice per week.
Yousef Yengej F.A., Jansen J., Ammerlaan C.M., Dilmen E., Pou Casellas C., Masereeuw R., Hoenderop J.G., Smeets B., Rookmaaker M.B., Verhaar M.C, & Clevers H. (2023). Tubuloid culture enables long-term expansion of functional human kidney tubule epithelium from iPSC-derived organoids. Proceedings of the National Academy of Sciences of the United States of America, 120(6), e2216836120.
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10

Evaluation of Uremic Toxins on Inflammation

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All reagents were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands) unless stated otherwise. In the present study, lipopolysaccharide (LPS) from Escherichia coli 0127:B8 was used. The toxins, p-cresyl glucuronide (pCG) and p-cresyl sulfate (pCS), were synthesized by the Institute for Molecules and Materials, Radboud University, Nijmegen, The Netherlands, as previously described [36 (link)]. IκB kinase (IKK) inhibitor BMS-345541 was obtained from Axon Medchem (Groningen, The Netherlands). Water (LC-MS grade), acetonitrile (ACN; HPLC-S grade) and methanol (HPLC grade) were obtained from Biosolve (Valkenswaard, The Netherlands). Formic acid (analytical grade) was purchased at Merck (Darmstadt, Germany). Ultrapure water was produced by a Milli-Q® Advantage A10 Water Purification Systems (Merck, Schiphol-Rijk, The Netherlands). Cell culture plates were obtained from Greiner Bio-One (Monroe, NC, USA).
Mihajlovic M., Krebber M.M., Yang Y., Ahmed S., Lozovanu V., Andreeva D., Verhaar M.C, & Masereeuw R. (2021). Protein-Bound Uremic Toxins Induce Reactive Oxygen Species-Dependent and Inflammasome-Mediated IL-1β Production in Kidney Proximal Tubule Cells. Biomedicines, 9(10), 1326.
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