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Objective : To investigate the cartilage cells and bone marrow mesenchymal stem cells (BMSCs) mixed culture for cartilage tissue engineering influence and determine the optimal ratio between the two . Method : Take a six -month-old New Zealand rabbits , male or female . Isolated and cultured in vitro amplification of rabbit articular chondrocytes and BMSCs. The first generation of the end of the harvest chondrocytes and BMSCs, in different proportions ( cartilage cells / BMSCs: 4/1, 2/1, 1/1, 1/2, 1/4, pure chondrocytes ) generation of mixed culture . To 4 × 107/ml final concentration of cells inoculated with 40μl polylactic acid polyglycolic acid polymer scaffold (PLGA: 4mm × 4mm × 2mm) cultured for 2 days on a static , cyclic pressure field into cultured ( pressure : 0 ~ 200kPa, frequency : 0.1 Hz, time : 8h / d) for three weeks. After three weeks of each group the size of tissue engineered cartilage morphology ﹑ ﹑ ﹑ luster and texture of tissue elasticity ; HE staining of tissue engineered cartilage cell proliferation and distribution ; toluidine blue staining amino glycosaminoglycan (GAG) secretion and distribution , and use of 1,9 dimethyl methylene blue assay for quantification of GAG content ; using collagen type Ⅱ immunohistochemical observation of type Ⅱ collagen secretion and distribution , and use image-proplus6.0 image analysis system type Ⅱ collagen staining area OK semi-quantitative analysis ; colorimetric determination of DNA content of tissue engineered cartilage tissue engineering cartilage observed cell proliferation . Results: The mixed culture of chondrocytes and BMSCs group compared with the simple chondrocytes , larger , smooth , elastic and shiny. Histology showed a dense mixed- culture group structure , more uniform distribution of the extracellular matrix , of which 2 /1 group showed lacuna . Mixed culture group Ⅱ collagen staining area , GAG content , DNA content higher than that of chondrocytes group , of which 2 /1 group was the highest , the difference was statistically significant (P lt; 0.05). Conclusion: mixed culture of chondrocytes and BMSCs can significantly improve the quality of tissue engineered cartilage , chondrocytes and BMSCs in which the concentration ratio of 2/ 1, the best results.
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