Cell viability of Panc-1 spheroids was measured using a 3D cell viability assay (Promega, Milan, Italy). Briefly, homogeneous spheroids were removed from the 96-well low-attachment culture plate and placed separately in single wells of a 96-well opaque culture plate (BD Falcon). CellTiter-Glo® 3D reagent was added to each well and the luminescence signal was read after 30 min with the GloMax® bioluminescent reader (Promega).
3d cell viability assay
Manufactured by Promega
Sourced in United States
The 3D Cell Viability Assay is a lab equipment product designed to measure the viability of cells in a three-dimensional (3D) culture system. It provides a quantitative assessment of cell health and proliferation in a 3D environment.
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Lab products found in correlation
Glomax bioluminescent reader, by Promega (1 mentions)
96 well opaque culture plate, by BD (1 mentions)
Celltiter glo 2d, by Promega (1 mentions)
Caspase glo 3 7 assay, by Promega (1 mentions)
Cck 8, by Promega (1 mentions)
Arvo x3, by PerkinElmer (1 mentions)
Foxm1, by Cell Signaling Technology (1 mentions)
Phospho histone h2ax γh2ax, by Cell Signaling Technology (1 mentions)
γ tubulin, by Santa Cruz Biotechnology (1 mentions)
β catenin, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Glutamine, by Lonza (1 mentions)
5 protocols using 3d cell viability assay
1
Measuring 3D Cell Viability of Panc-1 Spheroids
2
Anoikis-recovery and Cell Viability Assays
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Caspase-3 activity, cell viability and anoikis-recovery assays were performed using Caspase-Glo 3/7 Assay, CellTiter-Glo 2D or 3D Cell Viability Assay (Promega, Madison, WI) as described in detail in the Supplementary Methods . In the anoikis-recovery assay, cells were cultured in ultra-low attachment plates for 24 h before they were re-plated to the collagen I-coated 96-well plates to recover for 48 h.
3
Cell Viability Evaluation of Anticancer Treatments
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Two-dimensional and 3D cell viabilities were assessed using a CCK-8 and 3D cell viability assay (Promega, Madison, WI, USA), respectively. Two-dimensional cultures (8000 cells per well) and 3D cultures (one cell spheroid per well) were seeded in the 96-well plate and treated with various concentrations of gefitinib, rapamycin, gefitinib + rapamycin, NP, NP-Apt, NP (gefitinib + rapamycin), and NP-Apt (gefitinib + rapamycin) for 48 h. Next, CCK-8 solution and 3D cell viability assay reagent were added to each well, respectively. For 2D cultures, absorbance was measured at 450 nm (Thermo, Waltham, MA, USA) after 1 h of incubation at 37 °C. For 3D cultures, the luminescence signal (Thermo, USA) was read after 30 min of incubation at room temperature.
4
Cytotoxicity Assay of Anti-B7-H3 ADCs
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Cells were seeded into 96-well low-cell-adhesion cell culture plates at 2,000 cells per well. After overnight incubation, each serially diluted test substance (DS-7300a, parental anti-B7-H3 Ab, isotype control ADC, and DXd) was added then all of the plates were incubated for 6 days. After the incubation, cell viability was evaluated using 3D Cell Viability Assay (Promega Corp.) in accordance with the manufacturer's instructions. The luminescence was measured by a microplate reader, ARVO X3 (PerkinElmer Japan Co., Ltd.). The concentration of each substance inducing 50% inhibition (IC50) in each cell line was estimated in accordance with the sigmoid Emax model.
5
Stem Cell Viability Evaluation Protocol
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All media, sera, antibiotics, and glutamine for cell culture were from Lonza (Basel, Switzerland). Primary antibodies for western blotting were used according to the manufacturer’s protocol: β-Actin (#8227), poly-(ADPribose)-Polymerase (PARP)-Ab (#556494), phospho-Histone H2AX (γH2AX) (#05636), FOXM1 (#5436S), β-catenin (#8480S), Oct-4 (#2750S), were purchased from Cell signaling Technology (Danvers, MA, USA). γ-Tubulin (#sc-7396) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Secondary antibodies were purchased as follows: polyclonal goat anti-rabbit IgG (H + L)-HRP conjugate (#1706515) and polyclonal goat anti-mouse IgG (H + L)-HRP conjugate (#1706516) were purchased from Abcam (Cambridge, UK); polyclonal rabbit anti-goat IgG-HRP conjugate (#sc-2768) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Goat polyclonal Secondary Antibody to Mouse IgG - H&L - Alexa Fluor® 594 (#ab150120). Stem cell viability was evaluated by 3D Cell Viability Assay (Promega, Madison, WI, USA) according to the manufacturer’s protocol.
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