The Experimental Research Of Residual Ear Chondrocytes Of Microtia Proliferation In Vitro

Objective To investigate the changes of biological characteristics of residual ear chondrocytes proliferation in vitro, and study the effects of cryopreservation and CDMP-1 to proliferation and differentiation of the chondrocytes .Methods To collect 10 specimen residual ear cartilage of microtia patients about 8-year-old and cultured in vitro, dye with toluidine blue and immunohistochemistry of collagen type II. By the methods of the staining to study dedifferentiation of the chondrocytes subcultured in vitro and effects of CDMP-1 to differentiation of the chondrocytes. To investigate proliferation effects of CDMP-1 to the chondrocytes by MTT method . Analysis cell cycle of the chondrocytes after cryopreservation and subcultured with CDMP-1 on flow cytometric. Results The subcultured chondrocytes of residual ear lost their phenotype and show type II collagen stained cell and metachromatic GAG decrease. The positive type II collagen stained cell of 1st passage was about 71%, 3rd passage about 68%, 4th passage about 51% and 5th passage about 23%. After cryopreservation, the positive cell of 3rd one about 68%, like the same passage chondrocytes. The medium presenced with lOOng/ml CDMP-1 can maintain the positive chondrocytes of 4th passage to about 66%. Nither CDMP-1, nor cryopreservation can effect cell cycle of the chondrocytes. CDMP-1 can proliferate the residual ear chondrocytes by dose-effects, and the most efficient density is lOOng/ml. Conclusion The residual ear chondrocytes...