TABLE I.
Utilization of immortalized cell lines for spheroid formation.
| Cell type | Culture method | Spheroid Size | Remarks | Ref. |
|---|---|---|---|---|
| Human dermal fibroblasts | Rotational culture | 240 μm | Aggregates were inoculated on a scaffold of polyglycolic acid after the formation and could develop a new type of tissue engineered skin. | 41 |
| Lung adenocarcinoma (H358 and A549 cells) | 96-well plates coated with type I rat tail collagen | 60 μm | The 3D spheroids were more resistant to treatment with higher IC50 values for A549 and H358 cell lines compared to 2D. | 42 |
| MCF7 human breast cell | Encapsulation of cells in PEG-fibrinogen hydrogel microspheres made via hanging droplet | 100–400 μm | Cells encapsulated in gel-based microspheres had higher nuclear masses, a greater degree of disorganization, and enhanced tumorigenic morphology compared to those in spheroids. | 43 |
| Human fibroblasts, endothelial cells, and colon cell line (CRC and CCD841 CoN) | Low attachment 96-well U-bottom plate | 350–400 μm | Variations in drug combination efficacy between the cell types, cell ratios, and culture systems were observed. | 44 |
| Primary human hepatocytes (PHH) and Kupffer cells (KCs) | 96-well spheroid plates | 300– 350 μm | Inflammatory responses were recapitulated in co-culture spheroids, and 3D PHH spheroids with repeated dosing were more sensitive than 2D monolayer. | 45 |
| Normal human keratinocytes (NHKs) | 96-well round-bottom plate coated with a polymerized mixture of agarose (1.4%) and KSFM-scm | N/A | Spheroid-derived NHKs were enriched for a P63/K14 double-positive population that formed holoclonal colonies and reassembled into multicellular spheroids during 3D suspension subculture. | 46 |
| Ovarian cancer cells (OV-MZ-6, SKOV-3) | PEG-based hydrogels | > 50 μm | Spheroid formation was observed exclusively in 3D when cells were embedded within hydrogels. Proliferation in 3D was dependent on cell-integrin engagement and the ability of cells to proteolytically remodel their extracellular microenvironment. | 47 |
| LNCaP prostate cancer (PCa) cells | Hyaluronic acid (HA)–based bilayer hydrogel | 85 μm | HA-based bilayer platform supported the growth of prostate tumoroids, modeled paracrine interactions in the tumor microenvironment, and led to the production of pro-angiogenic signals in growing tumoroids. | 48 |