Adams S L, Pallante K M, Niu Z, Leboy P S, Golden E B, Pacifici M
Department of Anatomy and Histology, School of Dental Medicine, University of Pennsylvania, Philadelphia 19104-6003.
Exp Cell Res. 1991 Mar;193(1):190-7. doi: 10.1016/0014-4827(91)90555-9.
During endochondral ossification, small rapidly proliferating chondrocytes mature into flattened disc-shaped cells and then into large round hypertrophic cells. These morphological changes are accompanied by a decrease in the rate of cell proliferation. Type X collagen synthesis is initiated during chondrocyte maturation and reaches very high levels in the hypertrophic cells. We have analyzed type X collagen gene expression in chick embryo vertebral chondrocytes that were allowed to mature in monolayer culture and were then switched to suspension culture. The resuspended chondrocytes changed in shape from flat to round and decreased the proliferation rate as they do in vivo. These events were accompanied by a rapid, dramatic increase in type X collagen gene expression at the levels of transcription, steady-state mRNA and protein synthesis, as well as an increase in the number of cells producing type X collagen. The amount of type X collagen gene expression in resuspended chondrocytes was comparable to that in mineralizing cartilage in vivo. These results indicate that events accompanying the switch from monolayer to suspension culture (for example, the change from a flat to a round shape and/or the decrease in proliferation rate) may play a role in the induction of type X collagen gene expression during chondrocyte maturation. Thus we have developed an in vitro system that appears to mimic the events occurring during in vivo chondrocyte maturation. This in vitro model may provide an ideal system for further examination of the parameters regulating chondrocyte maturation and type X collagen gene expression.
在软骨内骨化过程中,快速增殖的小软骨细胞成熟为扁平的盘状细胞,然后成为大的圆形肥大细胞。这些形态学变化伴随着细胞增殖速率的降低。X型胶原合成在软骨细胞成熟过程中开始,并在肥大细胞中达到非常高的水平。我们分析了鸡胚椎体软骨细胞中X型胶原基因的表达,这些细胞在单层培养中成熟,然后转换为悬浮培养。重新悬浮的软骨细胞形状从扁平变为圆形,并且其增殖速率降低,如同在体内一样。这些变化伴随着X型胶原基因在转录、稳态mRNA和蛋白质合成水平上的快速、显著增加,以及产生X型胶原的细胞数量增加。重新悬浮的软骨细胞中X型胶原基因表达量与体内矿化软骨中的相当。这些结果表明,从单层培养转换为悬浮培养所伴随的事件(例如,从扁平到圆形的形状变化和/或增殖速率降低)可能在软骨细胞成熟过程中诱导X型胶原基因表达中发挥作用。因此,我们开发了一种体外系统,该系统似乎模拟了体内软骨细胞成熟过程中发生的事件。这种体外模型可能为进一步研究调节软骨细胞成熟和X型胶原基因表达的参数提供理想的系统。