Thyberg J, Hinek A
Cell Tissue Res. 1977 May 31;180(3):341-56. doi: 10.1007/BF00227600.
Chondrocytes were isolated enzymatically from rabbit ear cartilage, grown in vitro or as autotransplants for 1, 2 or 5 weeks and then examined by transmission electron microscopy. A confluent monolayer formed rapidly in vitro and the cells later grew in multiple overlapping layers, producing thick sheets of cartilaginous tissue. The cells retained a normal structure throughout the period of observation and, like the chondrocytes in intact cartilage, showed numerous microfilaments, and extensive granular endoplasmic reticulum and a prominent Golgi complex. Large amounts of intercellular matrix were laid down in vitro consisting of thin collagen fibrils, small rounded or polygonal granules believed to represent proteoglycans and patches and fibres of elastin. Chondrocytes in intramuscular autotransplants reconstituted an elastic cartilage. The exogenous origin of the cells in the transplants was verified by labeling of the lysosomes by exposure of the cells to colloidal thorium dioxide particles prior to injection. Structurally, the cells and the matrix of the transplants conformed to the above description. Accumulations of elastin-like material were sometimes observed in the Golgi vacuoles of the cells. Extracellularly, such conglomerates aggregated in connection with bundles of microfibrils, building up mature elastic fibres with a dense amorphous structure. The culture and transplant systems characterized here provide suitable experimental models for studies on development, growth and aging of elastic cartilage, including various aspects of the formation and turnover of elastic fibres and other macromolecular matrix components.
软骨细胞通过酶解法从兔耳软骨中分离出来,在体外培养或作为自体移植培养1、2或5周,然后用透射电子显微镜进行检查。在体外迅速形成汇合的单层细胞,随后细胞生长形成多个重叠层,产生厚的软骨组织片。在整个观察期间,细胞保持正常结构,并且与完整软骨中的软骨细胞一样,显示出大量微丝、广泛的颗粒内质网和突出的高尔基体复合体。在体外沉积了大量细胞间基质,其由细胶原纤维、被认为代表蛋白聚糖的小圆形或多边形颗粒以及弹性蛋白的斑块和纤维组成。肌肉内自体移植中的软骨细胞重建了弹性软骨。通过在注射前将细胞暴露于二氧化钍胶体颗粒来标记溶酶体,从而验证了移植中细胞的外源性起源。在结构上,移植的细胞和基质符合上述描述。有时在细胞的高尔基体泡中观察到类弹性蛋白物质的积累。在细胞外,这种聚集体与微原纤维束相连聚集,形成具有致密无定形结构的成熟弹性纤维。这里描述的培养和移植系统为研究弹性软骨的发育、生长和衰老提供了合适的实验模型,包括弹性纤维和其他大分子基质成分的形成和周转的各个方面。