Preparation of Cell-Paved and -Incorporated Polysaccharide Hollow Fibers Using a Microfluidic Device

Cellular constructs having hollow tubular structures are expected to be used as artificial blood vessels. We have recently demonstrated that water-insoluble polyion complexes (PICs) were formed from water-soluble polysaccharides with opposite charges at the interface of coaxial flows, which resulted in the formation of hollow fibers. In this study, both inside- and outside-cell-laden chondroitin sulfate C (CS)/chitosan (CHI) hollow fibers were prepared by utilizing a microfluidic device and modification with cell adhesive molecules. Loading of type I collagen (COL) and surface modification with fibronectin and gelatin using layer-by-layer assembly techniques improved the adhesion and spreading of fibroblast cells to/on the surface of CS/CHI hollow fibers. On the other hand, by suspending mesenchymal stem cells (MSCs) in the core flow solution, cells were successfully loaded in the walls of the hollow fibers. As the culture time extended, cells trapped in the PIC structures constituting the wall of the hollow fibers migrated to the interface between the hollow fibers and the medium: cells adhered to and stretched "on" the lumen surfaces in the COL-loaded fibers. In contrast, for the case of unmodified hollow fibers, it was difficult for cells to adhere to the lumen surfaces. Therefore, cell aggregates were formed "in" the lumen. Results of the live/dead assay and MTT assay clearly demonstrated that MSCs possessed certain levels of cell viability and proliferated for up to 10 days, especially for the cases of COL-loaded hollow fibers. On the basis of these results, the utility of the present hollow fibers in the formation of cellular constructs corresponding to blood vessels is also discussed.