组织培养法在关节软骨保存中应用.doc

组织培养法在关节软骨保存中应用 　 作者：宋洪强，亓建洪，玄燕华，毕研花 【摘要】 ［目的］探讨3种保存方法对关节软骨细胞活性的不同影响，寻求效果优良的软骨组织保存方法。［方法］切取成年猪骨软骨，制成约4.5 mm×5 mm大小的圆柱形骨软骨块。采用组织培养法、慢速梯度降温冷冻法、传统慢速连续降温冷冻法对软骨块进行保存处理，观察并比较保存后软骨细胞活性的变化。［结果］保存8周时，采用传统冷冻法的关节软骨细胞存活率不足50％，软骨基质成分大量丢失；采用慢速梯度降温冷冻法的细胞存活率66％，而使用组织培养法保存的关节软骨细胞存活率高达76%以上，软骨基质成分仅少量丢失。［结论］3种方法相比较，组织培养法可以长期保存关节软骨组织活性，是更为理想的软骨组织保存方法。 【关键词】 关节软骨； 保存； 组织培养； 活性 Abstract：［Objective］To study the effects of different storage method on the viability of cheodroeytes so as to find out the idem method for preservation of articular cartilage.［Method］Under the aseptic condition,acquiring 240 pieces of osteoehondral plugs from both condyles of femur and tibial plateau.The control group was not treatment using any preserved means.The tissuecultured group was to used F12DMEM medium longterm preserved osteochondral plugs under the conditions of 37 ℃.At preserve 8 weeks,takes the osteochondral section to carry on examination observation ohondrocytes Viability activeness change separately.［Result］The control group chnadrocyte survival rate was 94.4%.At preserves 8 weeks,the cell survival rate of the tissuecultured group was 76%,the gradingcooling cryopreservation group was 66%,the constantcooling cryopreservation group only was 41.80%.［Conclusion］Under the condition of 37 ℃,tissue culture method can long term preservation of vital articular cartilage in vitro.The chondrocyte survival rate of the preservation of vital articular cartilage in vitro.The chondrocyte survival rate of the tissuecultured method obvious higher than constantcooling cryopreservation method. Key words：articular cartilage; preservation; tissuecultured; viability 异体骨软骨移植物的保存尚无理想的方法，这明显制约了异体软骨移植的临床应用，如何保存软骨组织的活性，对于保证移植的成功至关重要。因此，作者进行了以下实验，希望找到更为理想的软骨组织保存方法。 1 材料与方法 1.1 主要试剂与仪器 F12-DMEM培养基，购于美国Sigma公司；程序降温仪器：国产RBLPAⅠ型程序冷冻仪；冷冻保存仪器：SANYO-80 ℃深低温冰箱。 1.2 实验动物 取清洁级健康成年长枫白猪10只20膝，无菌条件下在髌股关节及胫股关节切取正常骨软骨，制成直径约4.5 mm、长约5 mm的圆柱形骨软骨块240枚。将骨软骨块随机分为4组，每组60枚。备用。 1.3 实验方法 将骨软骨块随机分为4组，每组60枚。分别为： 对照组：即新鲜软骨标本不采取