Abstract:Objective To clarify the feasibility of PLLA and Cultispher G porous microspheres as an in vitro culture material for auricular chondrocytes,and to find a suitable microsphere scaffold for the three-dimensional dynamic culture of chondrocytes.Methods Aricular chondrocytes were isolated and purified from swine ear.P1 chondrocytes were inoculated on PLLA microspheres and Cultispher G microspheres,then cultured in a rotary cell culture system(RCCS).Growth of chondrocytes was observed on the microcarriers by using inverted and scanning electron microscopes.Proliferation rate of chondrocyte was measured by CCK-8 examination method.Relevant genes were measured by real-time PCR.Results Inverted and scanning electron microscopes showed chondrocytes could grow on PLLA microspheres and Cultispher G microspheres.Inoculation rates of chondrocytes were(37.67%±2.33%)and(73.67%±3.53%)respectively.Increasing fold of chondrocytes on PLLA microspheres was higher than that on Cultispher G microspheres after cultured for 21 days.Real-time PCR showed that PLLA microspheres maintained chondrocyte phenotype better than Cultispher G microspheres.Conclusion Compared with Cultispher G microsphere,PLLA porous microspheres are more suitable for three-dimensional culture of chondrocytes in vitro.

Key words:L-polylactide,Porous microsphere,Three-dimensional cultivation