Nunclon Sphera 3D culture system
Thermo Scientific Nunclon Sphera culture system is designed for spheroid culture (e.g., 3D tumor spheroids) and organoid culture (e.g., brain organoids) with a wide selection of format from plates to dishes and flasks. The 3D cell culture surface allows virtually no cell attachment to the cultureware, supporting the establishment of 3D spheroids and organoids via cell-to-cell aggregation through naturally secreted extracellular matrices (ECM).
Customer stories
I did a side-by-side comparison of the Nunclon Sphera and Corning® Costar® plates. In the Sphera plates, almost all cells aggregated nicely, whereas the Costar® plates, some cells were left out. Further, the spheroids in the Sphera plates created more pronounced protrusions/buds. I interpret this as Sphera plates having less adhesion, allowing the spheroids to change shape more freely. Since plate surface quality is a key parameter, I am using Sphera plates for my experiments.
— Keisuke Ishihara, Postdoctoral Researcher at the Max Planck Institute for Molecular Cell Biology and Genetics
Out of the different low attachment plates I have used to form 3D clusters of stem-cell derived pre-endocrine cells, Nunclon Sphera plates have been by far the best in terms of higher cell aggregation and lower cell death.
— Alberto Bartolom, PhD at Columbia University
It was much easier to maintain my spheroids in the Nunclon Sphera U-bottom plates than Corning ULA plates, as they were kept for a long time (21 days). The Sphera plates made it easier to administer dosages and sample from the round bottom. Moving forward, I will use the Sphera round-bottom plates for culturing spheroids.
— Carlemi Calitz, PhD at Postdoctoral Research Fellow at Uppsala University
In order to control the size of spheroids, we started using Nunclon Sphera and found out that the plate enabled to fabricate self-organized and size-controlled spheroids using mouse pre-osteoblast cells. We examined gene expression changes in the spheroids compared to the conventional 2D monolayer condition, resulted in up-regulations of osteocyte markers after 2 days of incubation in Nunclon Sphera. We thought that the cell condensation condition achieved in the spheroids triggered the osteocyte differentiation of pre-osteoblast cells. (J. Kim & T. Adachi (2020). J. Biomech. Sci. Eng., 15(3), 20-00227)”
— Dr. Jeonghyun Kim, Biomechanics Laboratory, Institute for Frontier Life and Medical Sciences, Kyoto University
Low cell attachment surface
Proprietary surface coating of Nunclon Sphera prohibits any protein adsorption to the cultureware surface, thereby minimizing monolayer cell adhesion to the culture vessel.
Figure 1. Compared to the standard cell culture flask (e.g., Nunclon Delta), minimal cell attachment was observed on the Nunclon Sphera 3D culture flask across 3 different cell types.
Consistent 3D tumor spheroid formation
The reliable cell growth and the ease in controlling the spheroid size on the Nunclon Sphera 96-well U-bottom 3D cell culture plate are important for applications where consistency and reproducibility are valued. 3D tumor spheroids imitate the in vivo development of solid tumors with homogenous growth and, in the cases of sizable tumor, the hypoxic core.
Figure 2. Growth kinetics of HCT 116 colon cancer cells shows early spheroid formation in the Nunclon Sphera 3D cell culture plate, even with low initial seeding density of 100 cells/well.
Figure 3. Size of the HCT 116 3D tumor spheroids can be controlled by the initial seeding densities in the wells. HCT 116 colon cancer cells were cultured on the Nunclon Sphera 3D cell culture plate for 112 hours.
Figure 4. The hypoxic core in a HeLa tumor spheroid after 2 days of culture on the Nunclon Sphera 96-well U-bottom 3D cell culture plate with initial seeding density of 1000 cells/well. The 3D tumor spheroid was stained with Invitrogen Image-iT Red Hypoxia Probe and Invitrogen NucBlue Live ReadyProbes reagent on a Nunc glass bottom dish.
3D cell culture protocols and applications
Protocols for spheroid generation
To help guide you in your 3D cell culture, our R&D teams have developed suggested protocols for the generation of spheroids from the most commonly used cancer cell lines using Nunclon Sphera cell culture plastics and Gibco Media.
| Cell line | Cancer model | Suggested protocol |
|---|---|---|
| SW480 | Colorectal cancer | Thermo Fisher Scientific R&D protocol |
| SKOV-3 | Ovarian cancer | Thermo Fisher Scientific R&D protocol |
| T47D | Breast cancer | Thermo Fisher Scientific R&D protocol |
| MDA-MB-231 | Breast cancer | Thermo Fisher Scientific R&D protocol |
| HepG2 | Liver cancer | Thermo Fisher Scientific R&D protocol |
| LNCaP | Prostate cancer | Thermo Fisher Scientific R&D protocol |
| PC-3 | Prostate cancer | Thermo Fisher Scientific R&D protocol |
| PANC-1 | Pancreatic cancer | Thermo Fisher Scientific R&D protocol |
| Prostate cancer |
Neurosphere and brain organoid generation
Through endogenously secreted natural ECM, neural stem cells (NSC) aggregate with one another, leading to rapid formation of neurosphere. With specific protocols, consistent cell growth in neurosphere can lead to further differentiation to multi-cellular structures known as brain organoid with tissue-specific morphology and function.
Figure 5. Human iPSC-derived neurospheres were imaged and stained with DAPI after being cultured on the Nunclon Sphera 3D cell culture plate for a week.
Figure 6. Brain organoid cultured in the Nunclon Sphera 3D cell culture dish was immunostained and imaged using Invitrogen EVOS FL Auto 2 imaging system at 10x magnification.
Embryoid body formation
Embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC) form 3D aggregates, known as embryoid body (EB), held together by complex network of adhesion molecules. The paracrine signaling within an embryoid body is crucial to embryogenesis in respond to environmental cues that lead to the eventual differentiation of three germ layers.
Figure 8. The differentiation of a human EB to three germ layers on Nunc Lab Tek II Chamber Slides as shown by the expression of ectoderm marker beta-tubulin, endoderm marker alpha fetoprotein (AFP), and mesoderm marker smooth muscle actin (SMA). Counter-stained with DAPI.
Model system for high-throughput screening (HTS) in drug discovery
High failure rate of new drug development that incurs enormous cost for both pharmaceutic companies and consumers calls for better 3D cell models in the early stages of the drug development process. 3D cell spheroids and organoids, with uniform size and centered location in the wells of the Nunclon Sphera 3D cell culture microplates, present an attractive and more biologically relevant 3D cell model system for drug discovery and disease modeling where high-throughput screening on high-content screening platforms is often employed.
Figure 9. A brightfield image of consistent A549 human lung 3D tumor spheroids with uniform size and shape in the Nunclon Sphera 96-well U-bottom plate.
Figure 10. Niclosamide-induced mitochondrial membrane depolarization and apoptosis are demonstrated using A549 and HeLa 3D tumor spheroids on the Nunclon Sphera 96-well U-bottom plate. Tumor spheroids are stained with MitoTracker Orange and CellEvent Green Caspase 3/7.
Ordering information
Related products
Additional resources
3D Cell Culture & Analysis Handbook
3D eLearning Module
Learn about spheroid and organoid culture techniques, applications, and various imaging modalities including high-content analysis through this interactive online course
Brochures
Application notes
Posters
For Research Use Only. Not for use in diagnostic procedures.
