Abstract:

BACKGROUND: Solid organ transplantation has been actively developed with a good therapeutic effect, but the lack of organ donor source is a worldwide problem.
OBJECTIVE: To develop a centrifugal cell seeded method for rapid and efficient tissue engineering construction.
METHODS: Different acceleration forces and cell densities and centrifugal times were studied. The orthogonal design method was used to optimize centrifugation parameters for cell seeding. To test stability and reproducibility of the orthogonal design results, experiments were studied using optimization centrifugation parameters. Monolayer cell adherence ratio after centrifugation and harvested cell numbers after cultured for 3 days was counted. Epithelial stratification tissues were constructed in vitro. The cell sheets were constructed on transwell culture insert using centrifugation parameters. Pathological sections were cut and stained with hematoxylin and eosin for 7 consecutive days.
RESULTS AND CONCLUSION: The (93.75±9.58)% attached cells were achieved using optimal cell seeding density at 9×105 cells/cm2 with a centrifugation at 2 200 r/min for 4 minutes. Monolayer centrifugal seeded cell sheets showed better cell viability and proliferation compared to naturally seeded cell sheets. The one-layer to four-layer cell sheets were rapidly constructed and showed good viability. Different layers of scaffold-free cell sheets were rapidly constructed using optimal centrifugation procedure. It demonstrates that the centrifugation method is a good way for tissue engineering construction.