Dr. Valentina Fermi (Mannheim, DE), Hannah Helt (Mannheim, DE), Annamarija Raic (Mannheim, DE), Nadine Kaiser (Ludwigshafen, DE), Veronique Schwartz (Mannheim, DE)
Abstract text (incl. figure legends and references)
Question. In vitro cell culture systems play a crucial role in drug discovery, basic research and regenerative medicine. In this context, 3D in vitro model systems are becoming increasingly relevant due to their potential to resemble in vivo microenvironment more reliably than 2D cell cultures. 3D spheroids allow to faithfully reproduce cell-cell and cell-matrix interactions for more precise validation of cell-based assays.
Methods. faCellitate has developed a chemically defined, animal-free coating solution which can be easily applied to cell culture surfaces. Being completely inert, the BIOFLOAT coating prevents unspecific binding of cells and proteins, thereby providing an innovative platform for the generation of 3D spheroids. Primary cells and cancer cell lines were used to benchmark this surface modification and spheroid morphology, cell viability and functionality as well as drug sensitivity were analysed.
Results. BIOFLOAT coating enables the rapid, reliable and reproducible generation of a single perfectly round-shaped spheroid in each well of pre-coated plates (Fig.1). Reproducible 3D spheroids are forming on BIOFLOAT surfaces also with relevant primary cells. For instance, this was observed using primary human hepatocytes which are highly relevant models in toxicological studies (Fig.2).
Conclusions. The BIOFLOAT platform represents a reliable system to establish 3D model-based assays for high throughput applications in basic and applied research. The development of this technology is in line with the efforts to surpass animal models.
Figure 1: Rapid generation of perfectly round-shaped spheroids. 3,000 3T3 cells were seeded in each well of 384-well plates. The generation of 3D spheroids was monitored for 96 hours. Scale bar = 400 µm.
Figure 2: Reproducible spheroid formation with relevant primary cells. 2,500 human primary hepatocytes were seeded in each well of 96-well plates. Spheroid formation was monitored for 8 days. Scale bar = 100 µm.