Spheroid formation in GravityTRAP™ ULA Plates
The GravityTRAP™ Ultra-Low Attachment (ULA) Plate represents a simple, flexible, and automation-compatible platform for the generation, long-term cultivation, observation and testing of scaffold-free 3D microtissue spheroids in 96 well format. Each plate consists of a special non-adhesively coated 96-well sterile-packaged GravityTRAP™ (Tissue Re-aggregation and Assay Plate), and lid.
IMPORTANT
InSphero recommends GravityTRAP™ ULA Plates for the generation of spheroids using immortalized cell lines that are known to readily form microtissues, or as a starting point for investigating whether or not a cell type can form self-aggregating, scaffold-free spheroids. InSphero recommends our patented GravityPLUS® Hanging Drop System (ISP-06-001,ISP-06-010) if generating spheroids in more complex 3D cell culture scenarios, such as when using primary cells, cell lines that are resistant to self-aggregation, or when generating co-culture microtissues (e.g., tumor/stroma). In such cases, the GravityPLUS® Hanging Drop System provides the greatest opportunity for success.
Figure 1: Spheroid formation in the GravityTRAP™ ULA Plate begins with initial seeding of cells in suspension, followed by a brief spin to concentrate cells. Following microtissue maturation,the SureXchange™ ledge of the tapered well facilitates medium exchange and compound dosing without disturbing or losing the microtissue.
Figure 2: HCT-116 and Hey tumor spheroids at day 3 of culture following seeding in the GravityTRAP™ ULA Plate at various cell densitites, demonstrating consistent, spheroid shaped microtissue formation.
Figure 3: Average size (day 3) of HCT-116 and Hey tumor spheroids as assessed by volume using the Cell3iMager. Standard deviation of 6 replicates per cell density shown for each cell line.
Figure 4: Average ATP content of HCT-116 and Hey tumor spheroids as assessed by Promega CellTiter-Glo® assay. Standard deviation of 6 replicates per cell density shown for each cell line.