Generation of Oxygenating Fluorinated Methacrylamide Chitosan Microparticles to Increase Cell Survival and Function in Large Liver Spheroids

• Published in: ACS applied materials & interfaces Vol. 14; no. 4; pp. 4899 - 4913
• Main Authors: Mansouri, Mona, Beemer, Samantha, Kothapalli, Chandrasekhar R, Rhoades, Tyler, Fodor, Petru S, Das, Dola, Leipzig, Nic D
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 02.02.2022
• Subjects: Acrylamides - chemistry; Acrylamides - metabolism; Acrylamides - pharmacology; Biocompatible Materials - chemistry; Biocompatible Materials - metabolism; Biocompatible Materials - pharmacology; Biological and Medical Applications of Materials and Interfaces; Cell Line; Cell Survival - drug effects; Chitosan - chemistry; Chitosan - metabolism; Chitosan - pharmacology; Halogenation; Humans; Materials Testing; Oxygen - chemistry; Oxygen - metabolism; Particle Size; Spheroids, Cellular - drug effects; Spheroids, Cellular - metabolism; Surface Properties; fluorinated microparticles; 3D in vitro model; microfluidic device; co-culture; liver spheroid
• ISSN: 1944-8244; 1944-8252
• DOI: 10.1021/acsami.1c19962

Despite advances in the development of complex culture technologies, the utility, survival, and function of large 3D cell aggregates, or spheroids, are impeded by mass transport limitations. The incorporation of engineered microparticles into these cell aggregates offers a promising approach to increase spheroid integrity through the creation of extracellular spaces to improve mass transport. In this study, we describe the formation of uniform oxygenating fluorinated methacrylamide chitosan (MACF) microparticles via a T-shaped microfluidic device, which when incorporated into spheroids increased extracellular spacing and enhanced oxygen transport via perfluorocarbon substitutions. The addition of MACF microparticles into large liver cell spheroids supported the formation of stable and large spheroids (>500 μm in diameter) made of a heterogeneous population of immortalized human hepatoma (HepG2) and hepatic stellate cells (HSCs) (4 HepG2/1 HSC), especially at a 150:1 ratio of cells to microparticles. Further, as confirmed by the albumin, urea, and CYP3A4 secretion amounts into the culture media, biological functionality was maintained over 10 days due to the incorporation of MACF microparticles as compared to controls without microparticles. Importantly, we demonstrated the utility of fluorinated microparticles in reducing the number of hypoxic cells within the core regions of spheroids, while also promoting the diffusion of other small molecules in and out of these 3D in vitro models.