3D Biomatrix has partnered with Cayman Chemical to provide a cell viability kit with a protocol optimized for use with Hanging Drop Plate-derived spheroids. The Perfecta3D Cell Viability Kit assay is based on the enzymatic cleavage of the tertrazolium salt WST-1 to formazan by cellular mitochondrial dehydrogenases in viable cells. You can use this assay to monitor cell viability after drug or toxicant treatment of spheroids created in the Perfecta3D Hanging Drop Plate provided with the kit. Sample data, shown in Figure 1 below, obtained by treating HEK293 cells with Triton™ X-100. Finally, an assay specifically optimized for Hanging Drop Plate-derived 3D cultures!


Spheroid Assay
Figure 1: Effect of initial cell number on absorbance at 450 nm. HEK293 cells were loaded onto a 3D Cell Culture Plate at different initial densities and grown at 37°C in a CO2 incubator. Four days later, cells were treated with vehicle, 0.001% triton X-100, or 0.01% triton X-100 for one hour and then transferred to a round bottom plate as described in the kit instructions. WST-1 solution was added to each well. The cells were then incubated in a 37°C incubator for 3.5 hours and then absorbance was measured at 450 nm.

    about the author

    Nicky Slawny trained as a developmental biologist with an unabashed interest in the mechanisms of stem cell differentiation. Her Ph.D. focused on using mouse embryonic stem cells as a model for lineage segregation during gastrulation. Currently, she is the Applications Director at 3D Biomatrix in Ann Arbor, Michigan. She has 15 years of tissue culture and molecular biology experience. In her free time she is an active Divemaster for a local scuba shop.

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