In Vitro Growth of Human Keratinocytes and OraludCancer Cells into Microtissues: An Aerosol-BasedudMicroencapsulation Technique

摘要

Cells encapsulation is a micro-technology widely applied in cell and tissue research, tissueudtransplantation, and regenerative medicine. In this paper, we proposed a growth of microtissue modeludfor the human keratinocytes (HaCaT) cell line and an oral squamous cell carcinoma (OSCC) cell lineud(ORL-48) based on a simple aerosol microencapsulation technique. At an extrusion rate of 20 �L/minudand air flow rate of 0.3 L/min programmed in the aerosol system, HaCaT and ORL-48 cells in alginateudmicrocapsules were encapsulated in microcapsules with a diameter ranging from 200 to 300 �m.udBoth cell lines were successfully grown into microtissues in the microcapsules of alginate withinud16 days of culture. The microtissues were characterized by using a live/dead cell viability assay,udfield emission-scanning electron microscopy (FE-SEM), fluorescence staining, and cell re-platingudexperiments. The microtissues of both cell types were viable after being extracted from the alginateudmembrane using alginate lyase. However, the microtissues of HaCaT and ORL-48 demonstrateduddifferences in both nucleus size and morphology. The microtissues with re-associated cells inudspheroids are potentially useful as a cell model for pharmacological studies.