A multiscale, vertical-flow perfusion system with integrated porous microchambers for upgrading multicellular spheroid culture. Issue 9 (11th April 2023)

• Record Type: Journal Article
• Authors: Takagi, Mai; Yamada, Masumi; Utoh, Rie; Seki, Minoru
• Is Part Of: Lab on a chip. Volume 23:Issue 9(2023)
• Page Start: 2257
• Page End: 2267
• Publication Date: 2023-04-11
• Subjects: Miniature electronic equipment -- Periodicals; Combinatorial chemistry -- Periodicals; Biotechnology -- Periodicals; 543.0813
• DOI: 10.1039/d3lc00168g
• Languages: English
• ISSNs: 1473-0197

Abstract

Abstract : A multiscale microfluidic system was developed that connects porous PDMS microchambers and microfluidic channels to generate a vertical flow of medium for spheroid culture. Abstract : Spheroid formation assisted by microengineered chambers is a versatile approach for morphology-controlled three-dimensional (3D) cell cultivation with physiological relevance to human tissues. However, the limitation in diffusion-based oxygen/nutrient transport has been a critical issue for the densely packed cells in spheroids, preventing maximization of cellular functions and thus limiting their biomedical applications. Here, we have developed a multiscale microfluidic system for the perfusion culture of spheroids, in which porous microchambers, connected with microfluidic channels, were engineered. A newly developed process of centrifugation-assisted replica molding and salt-leaching enabled the formation of single micrometer-sized pores on the chamber surface and in the substrate. The porous configuration generates a vertical flow to directly supply the medium to the spheroids, while avoiding the formation of stagnant flow regions. We created seamlessly integrated, all PDMS/silicone-based microfluidic devices with an array of microchambers. Spheroids of human liver cells (HepG2 cells) were formed and cultured under vertical-flow perfusion, and the proliferation ability and liver cell-specific functions were compared with those of cells cultured in non-porous chambers with a horizontal flow. The presented system realizes both size-controlled formation of spheroids and direct medium supply, making it suitable as a precision cell culture platform for drug development, disease modelling, and regenerative medicine.