유중수 액적 기반의 규모 가변적 3차원 스페로이드 배양법에 관한 연구

Abstract

Providing an environment mimicking the in vivo conditions, three-dimensional (3D) culture serves as an ex vivo model capable of independently replicating the biological mechanisms of organisms, cultivating micro-tissues derived from cells or stem cells. Among various 3D culture methods, scaffold-free 3D culture distinguishes itself by inducing only cell-cell interactions, maximizing intercellular communication. Techniques such as hanging drop, cell floater, and spinner tank fall into this category, primarily utilized for culturing spheroids or generating embryoid bodies, the precursors of organoids. Additionally, the Water-in-oil droplet culture, to be introduced hereafter, can be considered a scaffold-free 3D culture model, allowing the cultivation of micro-tissues based on the self-aggregation capability of cells. Water-in-oil droplet culture employs silicon oil, characterized by excellent thermal and oxidative stability, as the outer medium for cell medium droplets. The high gas permeability of the oil enables smooth cell cultivation. Furthermore, the oil not only maintains the droplet's shape close to a sphere but also serves as a barrier against external contaminants, restricting evaporation and providing advantages for the cultivation of individual micro-tissues. In this study, the utility of water-in-oil droplet culture was validated by comparing the formation, morphology, and growth rate of spheroids over the cultivation period with conventional models. Additionally, the applicability of this culture method to a channel-free microfluidic system is discussed.