Application of cell-sheet seeding to improve seeding efficiency of monolayer cells on the surface of biomaterials

Background: Cell seeding technique is a fundamental component of the tissue engineering construction. Effective cell seeding can not only shorten the construction time but also improve the final outcome of tissue engineered substitutes.

Objective: We improved the seeding efficiency of monolayer cells on the surface of biomaterials in tissue engineering. Cell sheet technology was applied to seed monolayer cell sheet instead of concentrated cell suspension, which was termed cell-sheet seeding.

Methods: Equal quantities of endothelial cells were seeded on the surface of equally sized acellular bovine pericardia by cell-sheet seeding and cell suspension seeding respectively. After culturing cell-seeded pericardia for 24 hours, the effects of these 2 cell seeding methods were evaluated and compared.

Results: As compared to cell suspension seeding, a larger quantity of loading cells, higher tissue-plasminogen activator (t-PA) and prostacyclin (PGI-2) content, and higher homogeneity of cells distribution on the surface of the pericardia was achieved with cell-sheet seeding. Furthermore, cell-sheet seeding did not affect metabolism or increase endothelial cells damage.

Conclusion: Cell-sheet seeding is an effective and time-saving method of seeding monolayer cells on the surface of biomaterials. This novel seeding method is particularly suitable for building tissue engineered substitutes with monolayered cells distribution and 2-dimensional structure.