【摘要】 目的制备以软骨微粒脱细胞基质为原料的微载体,为体外大量培养软骨细胞提供实验依据。方法利用胰蛋白酶等试剂制备直径为0.100～0.154mm的软骨微粒脱细胞基质并寻找胰蛋白酶的最佳含量及时间。取绵羊的新鲜膝、肘关节软骨600g,随机分为5组,每组8例,用不同含量的胰蛋白酶作用2～36h。标本进行大体观察,苏木精-伊红、胶原染色,光镜及扫描电镜观察,密度、含水量测定。结果微粒软骨脱细胞基质微载体无抗原性成分存在,主要含有胶原,氨基葡聚糖等混合细胞外基质成分。微粒呈不规则形,表面粗糙。软骨微粒脱细胞基质干重密度为0.6547g/mL,湿重密度为1.0627g/mL,含水量为72%。胰蛋白酶的最佳含量为0.25g/L,最适作用时间为(18.3±1.6)h。结论以软骨微粒为原料的微载体具有天然细胞外基质成分,是一种体外培养软骨细胞的良好载体。更多还原

【Abstract】 AIM:To prepare microcarriers made from cartilage microparticle acellular tissue matrix so as to provide experimental bases for mass culture of chondrocytes in vitro. METHODS:Microcarriers made from cartilage microparticle acellular tissue matrix were prepared with trypsin and other reagents,the diameter of which was from 0.100 mm to 0.154 mm,and then the optimal level and time of typsin were explored.Fresh goats’ knee and elbow articular cartilage,weighing 600 g, were divided randomly into 5 groups,each containing 8 cases.They were affected by different concentration of trypsin for 2-36 hours.The specimens were taken every 2 hours,then stained with haematoxylin eosin,collagen,and observed grossly under the light and scanning electron microscope for the measurement of the density and the content of water. RESULTS:The main components of these microcarriers were collagen,glycosaminoglycan,etc.,and no components with antigenicity were found.The microparticles arranged irregularly and had tough surface.The dry density of them was 0.6547 g/mL and the wet was 1.062 7 g/mL,and the content of water was 72%.The best concentration of trypsin was 2.5 g/L at the time of (18.3±1.6) hours. CONCLUSION:The microcarriers made from cartilage microparticle acellular tissue matrix contain nature excellular matrix and are good carriers for culturing cartilage cells in vitro.更多还原