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Ultralow adherent 24 well plates

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The Ultralow adherent 24-well plates are a specialized lab equipment designed for cell culture applications. These plates feature a surface treatment that minimizes cell attachment, promoting the growth of cells in a suspension format. The plates provide a controlled environment for researchers to cultivate cells while maintaining their desired phenotypes.

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Lab products found in correlation

Basic fibroblast growth factor (bfgf), by Merck Group (5 mentions) Insulin, by Merck Group (4 mentions) Epidermal growth factor (egf), by BD (4 mentions) Methylcellulose based medium, by STEMCELL (2 mentions) Serum free medium, by STEMCELL (1 mentions) Collagen type 1, by Advanced BioMatrix (1 mentions) Human epidermal growth factor (hegf), by Merck Group (1 mentions) Oncostatin m, by Thermo Fisher Scientific (1 mentions) B27 supplement, by Thermo Fisher Scientific (1 mentions) Dmem f12, by Thermo Fisher Scientific (1 mentions) Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions) Platelet derived growth factor, by Thermo Fisher Scientific (1 mentions) Tryple express, by Thermo Fisher Scientific (1 mentions) Methylcellulose, by Merck Group (1 mentions) Mammocult medium, by STEMCELL (1 mentions) Mc based serum free medium, by STEMCELL (1 mentions)

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24-well plates (1673 citations) 24-well ultralow plates (3 citations) 24 wells (2 citations)

8 protocols using ultralow adherent 24 well plates

1

Tumor Sphere Formation Assay

Cited 1 time
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Tumor sphere assay was performed under non-adherent and serum-free conditions as previously described as stem cell-selective conditions37 (link). Briefly, 1 × 104 CNT-exposed LFs and 5 × 103 H460 or BEAS-2B/SWCNT cells (2:1 ratio) were resuspended in 0.8% methylcellulose (MC)-based serum-free medium (Stem Cell Technologies, Vancouver, Canada) supplemented with 20 ng/mL epidermal growth factor (BD Biosciences), basic fibroblast growth factor and 4 mg/mL insulin (Sigma). The cell suspension was plated in ultralow adherent 24-well plates (Corning, Corning, NY) and cultured for two weeks. Large (>50 μm) tumor spheres were quantified under a light microscope.
Luanpitpong S., Wang L., Castranova V., Dinu C.Z., Issaragrisil S., Chen Y.C, & Rojanasakul Y. (2016). Induction of cancer-associated fibroblast-like cells by carbon nanotubes dictates its tumorigenicity. Scientific Reports, 6, 39558.
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2

Tumor Sphere Assay for Stem Cells

Cited 2 times
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Tumor sphere assay was performed under non-adherent and serum-free conditions as previously described as stem cell-selective conditions.25 (link),46 (link) Briefly, cells were suspended in 0.8% methylcellulose-based medium (Stem Cell Technologies, Vancouver, Canada) supplemented with 20 ng/mL epidermal growth factor (BD Biosciences, Franklin Lakes, NJ), basic fibroblast growth factor and 4 mg/mL insulin (Sigma-Aldrich, St Louis, MO) and plated at 5×103 cells in ultralow adherent 24-well plates (Corning, Corning, NY). Cells were cultured for two or three weeks and visualized under a light microscope.
Luanpitpong S., Li J., Manke A., Brundage K., Ellis E., McLaughlin S.L., Angsutararux P., Chanthra N., Voronkova M., Chen Y.C., Wang L., Chanvorachote P., Pei M., Issaragrisil S, & Rojanasakul Y. (2015). SLUG is required for SOX9 stabilization and functions to promote cancer stem cells and metastasis in human lung carcinoma. Oncogene, 35(22), 2824-2833.
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3

Neurosphere Migration in Collagen Matrices

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1 x 104 cells were cultured in Neurobasal Medium A supplemented with B27, 0.5 mM L-glutamine, 20 ng/mL hEGF (Sigma-Aldrich, St. Louis, MO), 20 ng/mL bFGF (Sigma-Aldrich) in ultra-low adherent 24-well plates (Corning Life Sciences, Corning, NY) for 24 h to allow formation of neurospheres [35 ]. Neurospheres of 75-150 um were resuspended in type I collagen (2.17 mg/mL)/DMEM + 10% FBS and appropriate treatments and plated in a 96-well plate pre-coated with type I collagen (Advanced BioMatrix, San Diego, CA). Neurospheres were imaged immediately (Time 0) with an Olympus CKX41 fitted with a Qicam Fast 1394 camera and then again at 15 and 24 h. Absolute maximal radius of each neurosphere was measured using ImageJ software and this distance was used to determine the relative change in radius. Three independent experiments were performed.
Knubel K.H., Pernu B.M., Sufit A., Nelson S., Pierce A.M, & Keating A.K. (2014). MerTK inhibition is a novel therapeutic approach for glioblastoma multiforme. Oncotarget, 5(5), 1338-1351.
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4

Clonal Mesensphere Formation Protocol

Cited 2 times
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For clonal mesensphere formation, BMSCs were plated at clonal density (∼1,000 cells/cm2) in ultralow adherent 24-well plates (Corning) as previously described (Mendez-Ferrer et al., 2010 (link), Pinho et al., 2013 (link)). The growth medium contained 15% chicken embryo extract (Fisher Scientific), 0.1 mM β-mercaptoethanol, 1% non-essential amino acids, 1% N2 and 2% B27 supplements (Gibco), basic fibroblast growth factor, insulin-like growth factor-1, epidermal growth factor, platelet-derived growth factor, and oncostatin M (Peprotech) (20 ng/ml) in DMEM/F12 (1:1)/human endothelial (Gibco) (1:2). The cultures were maintained at 37°C in a 5% CO2 water-jacketed incubator and left untouched for 1 week to prevent cell aggregation in low-density cultures. Half of the medium was changed weekly. The mesensphere colonies were detached with trypsin, and the cells were mechanically dispersed and re-plated back into ultralow adherent plates with culture medium. Secondary mesenspheres were counted after 7 days in culture.
Zhang P., Xing C., Rhodes S.D., He Y., Deng K., Li Z., He F., Zhu C., Nguyen L., Zhou Y., Chen S., Mohammad K.S., Guise T.A., Abdel-Wahab O., Xu M., Wang Q.F, & Yang F.C. (2016). Loss of Asxl1 Alters Self-Renewal and Cell Fate of Bone Marrow Stromal Cell, Leading to Bohring-Opitz-like Syndrome in Mice. Stem Cell Reports, 6(6), 914-925.
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5

Quantifying Tumor Sphere Formation in Cancer Cell Lines

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HSC-3 or CAL27 cells were treated with 10 μM CH223191, 10 μM CB7993113, 0.5 μM FICZ, 1 nM TCDD, and/or vehicle (0.1% DMSO or PBS) every 24 hours. After 48 hours, cells were harvested, dosed, and 3 x 103 cells plated in MammoCult Medium (STEMCELL Technologies) containing 0.5 μg/ml hydrocortisone, 2 mM L-glutamine, 100 I.U. penicillin/100 μg/ml streptomycin, and 1% methylcellulose (Sigma-Aldrich) in ultra-low adherent 24-well plates (Corning). Colonies were quantified with a Celigo S Imaging Cytometer (Brooks Automation, Chelmsford, MA) after 8 days. For secondary sphere formation, tumorspheres were mechanically and enzymatically dissociated with TrypLE Express (Gibco) for 10 minutes at 37° C until a single cell suspension was formed. The cells were then re-plated and imaged as above.
Stanford E.A., Ramirez-Cardenas A., Wang Z., Novikov O., Alamoud K., Koutrakis P., Mizgerd J.P., Genco C.A., Kukuruzinska M.A., Monti S., Bais M.V, & Sherr D.H. (2016). Role for the Aryl Hydrocarbon Receptor and Diverse Ligands In Oral Squamous Cell Carcinoma Migration and Tumorigenesis. Molecular cancer research : MCR, 14(8), 696-706.
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6

Tumor Sphere Assay for Cancer Stemness

Cited 2 times
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Tumor sphere assay was performed under non-adherent and serum-free conditions as previously described as stem cell-selective conditions [31 (link),36 (link)]. Briefly, cells were resuspended in 0.8% methylcellulose (MC)-based serum-free medium (Stem Cell Technologies, Vancouver, Canada) supplemented with 20 ng/mL epidermal growth factor (BD Biosciences, San Jose, CA), basic fibroblast growth factor and 4 mg/mL insulin (Sigma) and plated at 5 × 103 cells (BC, BSW, and H460) or 1 × 104 cells (SAC and SASW) in ultralow adherent 24-well plates (Corning, Corning, NY). Cells were cultured for two or three weeks. In order to assess the self-renewing property of cells, spheres were collected by gentle centrifugation, dissociated into single cell suspensions, filtered and cultured under conditions described above (second spheres).
Luanpitpong S., Wang L., Castranova V, & Rojanasakul Y. (2014). Induction of stem-like cells with malignant properties by chronic exposure of human lung epithelial cells to single-walled carbon nanotubes. Particle and Fibre Toxicology, 11, 22.
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7

Tumor Sphere Assay for Stem Cells

Cited 2 times
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Tumor sphere assay was performed under non-adherent and serum-free conditions as previously described as stem cell-selective conditions.25 (link),46 (link) Briefly, cells were suspended in 0.8% methylcellulose-based medium (Stem Cell Technologies, Vancouver, Canada) supplemented with 20 ng/mL epidermal growth factor (BD Biosciences, Franklin Lakes, NJ), basic fibroblast growth factor and 4 mg/mL insulin (Sigma-Aldrich, St Louis, MO) and plated at 5×103 cells in ultralow adherent 24-well plates (Corning, Corning, NY). Cells were cultured for two or three weeks and visualized under a light microscope.
Luanpitpong S., Li J., Manke A., Brundage K., Ellis E., McLaughlin S.L., Angsutararux P., Chanthra N., Voronkova M., Chen Y.C., Wang L., Chanvorachote P., Pei M., Issaragrisil S, & Rojanasakul Y. (2015). SLUG is required for SOX9 stabilization and functions to promote cancer stem cells and metastasis in human lung carcinoma. Oncogene, 35(22), 2824-2833.
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8

Mammosphere Formation Assay

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Adherent cells were counted and 2,000–4,000 cells were seeded in ultra-low adherent 24-well plates (Corning). Mammosphere medium was DMEM/F12 with 2% B27, 20 ng/mL EGF, and 4 µg/mL insulin and growing mammospheres were supplemented with medium every 3 days. After 7–14 days the mammospheres were counted under a microscope and collected for downstream analysis.
Carpenter R.L., Sirkisoon S., Zhu D., Rimkus T., Harrison A., Anderson A., Paw I., Qasem S., Xing F., Liu Y., Chan M., Metheny-Barlow L., Pasche B.C., Debinski W., Watabe K, & Lo H.W. (2017). Combined inhibition of AKT and HSF1 suppresses breast cancer stem cells and tumor growth. Oncotarget, 8(43), 73947-73963.
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