The Biological Characteristics Of Chondrocytes Cultured In Monolayer In Vitro And Seeded Into Gelatin Sponge

Objective To investigate the biological properties of chondrocytes and those seeded into gelatin sponge at different passages for providing available seed-cells for tissue engineering. To evaluate the advantage of gelatin sponge as scaffold for three-dimensional chondroncytes culture. To imporove the culture method by observing the biological characteristics on different plate. Methods The two-week rabbit articular chondrocytes were isolated and cultured and subcultured to observe the variation of biological characteristics such as growth curves and morphological changes by immunohistochemical analysis and HE staining. Chondrocytes in different passages seeded into gel scaffold were observed using confocal microscopy and electron microscopy including scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Some chondrocytes were stored by using cryopreservation. Results (l)The growth curves showed that the proliferation of the primary to the fourth passage was powerful. (2) It is easier for the fourth passage (P/0 chondrocytes and gelatin sponge coming into being complex. (3) The poly-hedral angle-shaped chondrocytes gradually were changed into spindle-shaped cells after the fifth passage .(4)The time for subcultured chondrocytes to adhere plant disk was shorter than that for the chondrocytes in the first passage. (5)The ratio of adhesion on cover-glass with rat tailor gel is the largest of three culture plates. Conclusions (1) On the basis of the chondrocytic attachment , proliferation and the ability of collagen type- II synthesis , the fourth-passage (P4) chondrocytes are best for cartilage tissue engineering of all different passages. (2) Gelatin sponge demonstrates overall biocompatibility and appropriate mechanical properties, as well as promote tissue formation. Gelatin sponage is suit for cartilage tissue engineering as three-dimensionalcell carrier. (3) chondrocytes cultured on cover-glass with rat tail gel grows more and adhere quicker than those cultured on other plates..