Media additives to promote spheroid circularity and compactness in hanging drop platform

Abstract

Three-dimensional spheroid cultures have become increasingly popular as drug screening platforms, especially with the advent of different high throughput spheroid forming technologies. However, comparing drug efficacy across different cell types in spheroid culture can be difficult due to variations in spheroid morphologies and transport characteristics. Improving the reproducibility of compact, circular spheroids contributes to standardizing and increasing the fidelity of the desired gradient profiles in these drug screening three-dimensional tissue cultures. In this study we discuss the role that circularity and compaction has on spheroids, and demonstrate the impact methylcellulose (MethoCel) and collagen additives in the culture media can contribute to more compact and circular spheroid morphology. We demonstrate that improved spheroid formation is not a simple function of increased viscosity of the different macromolecule additives, suggesting that other macromolecular characteristics contribute to improved spheroid formation. Of the various macromolecular additives tested for hanging drop culture, MethoCel provided the most desirable spheroid formation. Additionally, the higher viscosity of MethoCel-containing media improved the ease of imaging of cellular spheroids within hanging drop cultures by reducing motion-induced image blur.

CCN6 regulates IGF2BP2 and HMGA2 signaling in metaplastic carcinomas of the breast

“…Two weeks after implantation, mice were treated with rhCCN6 (1ng/g, i.v., twice a week) or BSA (1ng/g) in the same regimen, and followed by palpation and caliper measurements for 3 weeks. Organoids derived form MMTV-Cre; Ccn6 fl/fl mice tumors were generated following previous reports [20]. …”

Formation of stable small cell number three-dimensional ovarian cancer spheroids using hanging drop arrays for preclinical drug sensitivity assays

“…These plates have been used to generate human epithelial carcinoma spheroids, prostate cancer spheroids as well as non-cancer primary stem- and progenitor cell spheroids [14, 15]. Recently, Leung et al demonstrated that the spheroids generated using hanging drop array platforms demonstrate excellent circularity and compactness [16]. …”

Dispersible oxygen microsensors map oxygen gradients in three-dimensional cell cultures

“…The MDA-MB 231 cells used for this portion of the study do not spontaneously form tight spheroids 22 , enabling us to simulate a 2D slice of a spheroid in a 3D culture-like environment. Employing a prevalent practice for the qualification of bioreactors 35 , we generated a computational model of oxygen supply, diffusion, and uptake (Figure 3) by using the geometry of the experimental system and a range of literature values of oxygen V max and K m 36–42 to run a parametric sweep.…”

“…Spheroids were generated using high-throughput hanging-drop-array plates 21 and the culture methods recently presented 22 . Prior to use, a hydrophilic coating was applied onto the entire surface of the hanging-drop plate by soaking the plate overnight in 0.1%, Pluronic F108.…”