Isolation Of Rabbit Articular Chondrocytes Culte And Identification

Objective:Optimize the rabbit articular cartilage cells separation method,to explore the single-layer training process conditions,Telomerase and biological function characteristics.Methods:Choose 4 weeks, health,New Zealand male big white articular cartilage,adopting mechanical,enzyme digestion method for the separation of articular cartilage cells obtained.Determination of survival and cell in the womb of bovine serum containing 10 percent DMEM high sugar for in vitro culture in training.Stay cell fusion rate 85 percent to 90 percent by trypsin digestion,collect and with the same density stamped nestin.Generational observation and application of cellular MTT method drawing cell growth curve,While toluidine blue stain and RT-PCR method of dynamic study each generation of articular cartilage cells of I, II proteoglycan, type,collagen and telomerase gene expression changes the level.Results:This method rabbit articular cartilage cells is simple, efficient and collection process per gram cartilage gets 2 x 106 cells.In the process of training in vitro, with the number of chondrocytes nestin increases, the longer time,cell nestin form to"fiber cell" transform-ation.type collagen genes expression level of increasing trend, II type collagen and telomerase genes expression level of increasing trend, extracellular matrix polysa-ccharide decrease.Three former high cell biology and functional characteristics,mainly for I after generations of collagen, the high exp-ression of cartilage cells of collagen and II proteoglycan almost disappear.The trend appears to differentiation. Conclusions:1 The articular cartilage cells machinery, Double enzyme access method is simple and easy to operate,and can obtain a higher survival rate of cartilage cells.2 The rabbit in vitro single-layer training before the articular cartilage cells, biology and three phenotype stable function characteristic expression.