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24 well ultra low adherent plate

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The 24-well ultra-low adherent plate is a laboratory tool designed to provide a non-adhesive cell culture surface. It is suitable for the growth and maintenance of cells that prefer a low-attachment environment, such as stem cells, spheroids, and suspension cultures. The plate is made of a specialized material that minimizes cellular attachment, allowing for the formation and study of three-dimensional cell structures.

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Lab products found in correlation

Mammocult proliferation supplement, by STEMCELL (4 mentions) Stem cell factor (scf), by STEMCELL (1 mentions) Epidermal growth factor (egf), by STEMCELL (1 mentions) Basic fibroblast growth factor (bfgf), by STEMCELL (1 mentions) Dulbecco modified eagle medium (dmem), by STEMCELL (1 mentions) Mammocult basal medium human, by STEMCELL (1 mentions) Insulin, by STEMCELL (1 mentions) Methocult medium, by STEMCELL (1 mentions) Hydrocortisone, by STEMCELL (1 mentions) Inverted fluorescence microscope, by Zeiss (1 mentions) Mammocult, by STEMCELL (1 mentions) Inverted fluorescence microscope, by Nikon (1 mentions) Stem cell medium, by Thermo Fisher Scientific (1 mentions) Axiovision software, by Zeiss (1 mentions) Axiovert 40, by Zeiss (1 mentions)

Spelling variants of this product

24-well plates (1673 citations) Ultra Low plates (3 citations) 24 wells (2 citations)

7 protocols using 24 well ultra low adherent plate

1

Tumor Sphere Formation Assay

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Sorted cells (1 × 103 cells) were seeded in a 24-well ultra-low adherent plate (Corning) in 0.5 mL of mixed medium to perform sphere formation detection. The medium contained 32% MethoCult medium, 20% MammoCult basal human medium with a final concentration of 2% MammoCult proliferation supplements and 48% DMEM supplemented with final concentrations of 100 pg/mL EGF, 50 ng/mL bFGF, 5 ng/mL stem cell factor, 1 μM hydrocortisone, and 5 mg/mL insulin, all obtained from STEMCELL Technologies. The cells were cultured at 37 °C in a 1% O2 and 5% CO2 humidified atmosphere for 14 d. The number of tumor spheres was counted under microscope.
Sun T., Yang W., Toprani S.M., Guo W., He L., DeLeo A.B., Ferrone S., Zhang G., Wang E., Lin Z., Hu P, & Wang X. (2020). Induction of immunogenic cell death in radiation-resistant breast cancer stem cells by repurposing anti-alcoholism drug disulfiram. Cell Communication and Signaling : CCS, 18, 36.
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2

Sphere Formation Assay for Stem Cell Culture

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Sphere formation was performed by seeding 600–1000 cells/well in a 24-well ultra-low adherent plate (Corning Incorporated, Corning, NY, USA) in 500 μL of mixed medium containing 32% MethoCult medium, 20% MammoCult basal human medium with a final concentration of 2% MammoCult proliferation supplements (STEMCELL Technologies) and 48% DMEM supplemented with final concentrations of 100 pg/mL EGF, 50 ng/mL bFGF, 5 ng/mL stem cell factor, 1 × 10−6 M hydrocortisone, and 5 μg/mL insulin. The cells were cultured at 37 °C in a 1% O2 and 5% CO2 humidified atmosphere for 18 days (SW1353 cells) and 11 days (CS-1 cells). Photographs were taken and spheres were quantified by counting sphere numbers/well.
Wang K., Michelakos T., Wang B., Shang Z., DeLeo A.B., Duan Z., Hornicek F.J., Schwab J.H, & Wang X. (2021). Targeting cancer stem cells by disulfiram and copper sensitizes radioresistant chondrosarcoma to radiation. Cancer letters, 505, 37-48.
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3

Sphere Formation Assay for Stem Cell Culture

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Sphere formation was performed by seeding 600–1000 cells/well in a 24-well ultra-low adherent plate (Corning Incorporated, Corning, NY, USA) in 500 μL of mixed medium containing 32% MethoCult medium, 20% MammoCult basal human medium with a final concentration of 2% MammoCult proliferation supplements (STEMCELL Technologies) and 48% DMEM supplemented with final concentrations of 100 pg/mL EGF, 50 ng/mL bFGF, 5 ng/mL stem cell factor, 1 × 10−6 M hydrocortisone, and 5 μg/mL insulin. The cells were cultured at 37 °C in a 1% O2 and 5% CO2 humidified atmosphere for 18 days (SW1353 cells) and 11 days (CS-1 cells). Photographs were taken and spheres were quantified by counting sphere numbers/well.
Wang K., Michelakos T., Wang B., Shang Z., DeLeo A.B., Duan Z., Hornicek F.J., Schwab J.H, & Wang X. (2021). Targeting cancer stem cells by disulfiram and copper sensitizes radioresistant chondrosarcoma to radiation. Cancer letters, 505, 37-48.
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4

Mammosphere Formation Assay

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Cells were plated in 6-well plates at a density of 2x105 cells/well in 2 mL of complete medium and received fractionated irradiation. Twenty-four hours after irradiation, cells were treated with DSF/Cu (2.5 μM/1 μM) or 1 μM NF-κB inhibitor IMD0354 for an additional 24 hours. Mammosphere formation was performed by seeding the cells (1000 cells/well) in a 24-well ultra-low adherent plate (Corning Incorporated) in 500 μL of mixed medium containing 32% MethoCult medium, 20% MammoCult basal human medium with a final concentration of 2% MammoCult proliferation supplements (STEMCELL Technologies), and 48% DMEM supplemented with final concentrations of 100 pg/mL EGF, 50 ng/mL bFGF, 5 ng/mL stem cell factor, 1×10−6 M hydrocortisone, and 5 μg/mL insulin. The cells were cultured at 37°C in a 5% O2 and 5% CO2 humidified atmosphere for 14 days. Mammosphere pictures were taken using a Zeiss Inverted Fluorescence Microscope on day 10 or 14. Numbers of mammosphere were count on day 14. Mammosphere formation was also performed by seeding (1000 cells/well) in a 24-well ultra-low adherent plate of the Fluorescence Activated Cell Sorting (FACS) sorted ALDHneg cells or cells from a single cell suspension collected from a disaggregated tumor.
Wang Y., Li W., Patel S.S., Cong J., Zhang N., Sabbatino F., Liu X., Qi Y., Huang P., Lee H., Taghian A., Li J.J., DeLeo A.B., Ferrone S., Epperly M.W., Ferrone C.R., Ly A., Brachtel E.F, & Wang X. (2014). Blocking the formation of radiation–induced breast cancer stem cells. Oncotarget, 5(11), 3743-3755.
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5

Tumor Cell Sphere Formation Assay

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The isolated macrophages or tumor cells were seeded into 24-well ultra-low adherent plates (Corning, USA) at 1000 cells/well in 1 ml SCM (Stem Cell Medium, Gibco, ThermoFisher Scientific, Waltham, MA, USA) for 7 days. The number of spheres with diameter ⩾70 μm was determined using an inverted fluorescence microscope (Nikon). Sphere formation efficiency (SFE) was calculated as the percentage of seeded cells that gave rise to a spheroid.
Zhang W., Han Q., Ding Y., Zhou H., Chen Z., Wang J., Xiang J., Song Z., Abbas M, & Shi L. (2022). Bcl6 drives stem-like memory macrophages differentiation to foster tumor progression. Cellular and Molecular Life Sciences, 80(1), 14.
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6

Sphere and Colony Formation Assay

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Cells were seeded at 60–70% confluency in 10cm plates and treated at indicated times as previously described. 500 cells were plated in triplicates in 24-well ultra-low adherent plates (Corning, cat #3473) with 1ml of stem cell medium (for spheroid formation assay) or 6-well plates in 2ml fresh complete media (for colony formation assay). Cells were allowed to grow for 14 days for spheroid formation or 5–7 days for colony formation. Spheroid size and morphology were assessed using a Zeiss Axiovert 40 inverted microscope with Axio-Vision software (Carl Zeiss MicroImaging). Colonies were stained with crystal violet (0.5%). Spheres or clusters smaller than 100 μm were not counted.
Wang W., Fang F., Ozes A, & Nephew K.P. (2021). Targeting Ovarian Cancer Stem Cells by Dual Inhibition of HOTAIR and DNA Methylation. Molecular cancer therapeutics, 20(6), 1092-1101.
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7

Culturing Tumor-Derived Macrophages

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Macrophages isolated from tumors were seeded with DMEM into a 10 cm dish and incubated for 40 min. After washing away non-adherent cells, adherent cells were seeded into 24-well ultra-low adherent plates (Corning, USA) at 1000 cells/well in 1 ml SCM harboring 20 ng/ml M-CSF for 7 days. Then 1 ml DMEM containing 10% FBS and 1% penicillin streptomycin was added per 2 days for 5–6 weeks.
Zhang W., Han Q., Ding Y., Zhou H., Chen Z., Wang J., Xiang J., Song Z., Abbas M, & Shi L. (2022). Bcl6 drives stem-like memory macrophages differentiation to foster tumor progression. Cellular and Molecular Life Sciences, 80(1), 14.
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