Multicellular 3D spheroids can be generated by various methods, including the (1) spontaneous spheroid formation using (ultra) low binding plates; (2) “hanging drop” technique; (3) suspension cultures (e.g., by spinner flasks or bioreactors); (4) scaffold-based models (e.g., hydrogels), and (5) magnetic levitation [165]. Since spontaneous spheroid formation and the hanging drop method are arguably the most cost-effective and likely take the least amount of specialized skill set, we will focus on these two iterations of the model (Fig. 8). One of the pioneering approaches to generate self-assembling multicellular 3D tumor endothelial spheroids in hanging drops was by Timmins et al. [166]. Subsequently, this assay was validated by various groups and expanded on in terms of cell types tested and it evolved as an instrument to be transplanted into animals, devoid of Matrigel, to study cancer progression and ultimately metastasis
Multicellular 3D spheroids can be generated by various methods, including the (1) <br>spontaneous spheroid formation using (ultra) low binding plates; (2) “hanging drop” <br>technique; (3) suspension cultures (e.g., by spinner flasks or bioreactors); (4) scaffold-based <br>models (e.g., hydrogels), and (5) magnetic levitation [165]. Since spontaneous spheroid <br>formation and the hanging drop method are arguably the most cost-effective and likely take <br>the least amount of specialized skill set, we will focus on these two iterations of the model <br>(Fig. 8). One of the pioneering approaches to generate self-assembling multicellular 3D <br>tumor endothelial spheroids in hanging drops was by Timmins et al. [166]. Subsequently, <br>this assay was validated by various groups and expanded on in terms of cell types tested and it evolved as an instrument to be transplanted into animals, devoid of Matrigel, to study <br>cancer progression and ultimately metastasis
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