Table 1.
Comparison of 3D CSCs culture methods (Advantages and disadvantages)
| Method | Advantages | Disadvantages |
|---|---|---|
| Organoids | • The possibility of creating tumor-specific therapy | • The need to collect material from the patient |
| • They reflect tumor morphology and heterogeneity | • Establishment of specific cell ratio at the beginning | |
| • Relatively simple to maintain | • Selection of appropriate medium with cell growth factors or inhibitors | |
| • Ready-made protocols for various cancer types | • Lack of immune, muscle, nerve, and vascular cells to completely recreate tumor environment | |
| • The ability to derive high-throughput drug testing | ||
| • A good tool to investigate CSCs | ||
| • Organoids bio-bank-currently in development | ||
| Single cell derived spheroids | • Possible to detect CSC and distinguish subpopulations | • The necessity to obtain a single cell type by using FACS or limiting dilution protocol |
| • Possible to study only one type of cell reaction for a specific drug | • The optimal effect, while using advanced technology-microflow technique and special plates | |
| • Thanks to a microchip, the medium flow can be preserved and high-throughput drug screening is possible | ||
| Hanging drop | • One of the easiest method to obtain spheroids | • Might dry if the humidity is not well controlled |
| • Relatively cheap technique | • Time-consuming pipetting and medium change | |
| • The scaffold-free technique allows to avoid some cell-material interactions | • A small size of droplets and a small number of cells can be used | |
| ULA plate | • Ready for automated method and high-throughput research | • Lack of microenvironment reflection |
| • Wide range of cell quantity used for spheroid formation | • Might be difficult in drug distribution and imaging because of high compact spheroid | |
| • Homogenous spheroids with similar morphology | • Possible damage of formed spheroid during medium changing | |
| Hydrogels and polymers | • Opportunity to adjust pore size and stiffness of a drop | • Different results for different hydrogels/polymers |
| • A low-cost and efficient platform for spheroid forming | • Might interact with cells | |
| • Stable and biodegradable form | • CSCs markers expression depends on drop stiffness | |
| • Modification of a hydrogel or polymer composition might change gene expression | ||
| Solid-phase spheroids | • Easy to maintain 3D culture | • Some hydrophobic powder might be cytotoxic |
| • No need for supporting scaffold (only liquid drop is covered by hydrophobic powder) | • Difficulties in real-time imaging | |
| • Low-cost technique | • Might be difficult to provide medium exchange without structural damage |