Influence of alginate-based microcapsule permeability on 3D cancer cell cluster proliferation

Background: Functional interaction between cancer cells and the surrounding microenvironment is still not sufficiently understood, which motivates the tremendous interest in the development of numerous in vitro and in vivo tumor models. Study Aims: To study the influence of different permeabilities of microcapsules (MCs) on different cancer cell proliferations, and to design and engineer the formation of 3D tumor clusters in MCs. Materials and Methods: A fluidics-based low-cost methodology was used to reproducibly generate alginate (AL) and alginate-chitosan (AL-CS) MCs in a cross-junctions water-in-oil system. The diffusion through the shell of AL and AL-CS MCs was monitored using fluorescein sodium (376 Da), FITC-Dextran 70 (70 kDa), and FITC-Dextran 2000 (2000 kDa) as fluorescent probes representing small molecules, proteins, and macromolecules, respectively. HepG2 Red FLuc (human hepatoma cell line) and A375 (human melanoma cell line) cultured in high-glucose DMEM medium were used to study the proliferation differences in terms of dimensions and geometries in AL and AL-CS MCs. The metabolic activity of tumor clusters in MCs was confirmed by tracking the turnover of testosterone to androstenedione with lower case Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). Results: HepG2 Red FLuc and A375 cells show different proliferation properties in AL and AL-CS MCs. A375 tumor clusters grow faster in more permeable AL MCs and slower in less permeable AL-CS MCs. In the case of HepG2 Red FLuc, a significant difference in proliferation rate was not observed between AL and AL-CS MCs at the early stage (1 week). Interestingly, it was observed that different loose and tight cell cluster morphologies can form, also including cell proliferation along radial directions in both MC types and both cell lines. Cytochrome P450 (CYP)-dependent metabolization of testosterone by both HepG2 Red FLuc and A375 tumor clusters in the AL and AL-CS MCs showed the same trends in good