中国组织工程研究2017，Vol.21Issue(12)：1843-1848,6.DOI:10.3969/j.issn.2095-4344.2017.12.007
软骨细胞膜片的成软骨性能
Chondrogenesis of chondrocyte cell sheets
摘要
Abstract
BACKGROUND:The cell-sheet technology, based on a temperature-responsive culture, has been drawing more and more attention;however, the temperature-responsive culture dish is quite expensive. Therefore, it is imperative to develop a substitutive technique.OBJECTIVE: To study the feasibility of cell-sheet ulturing using common culture dish, and investigate the chondrogenesis of the cell sheet. METHODS: A piece of nasal septal cartilage was adopted from a patient with deviation of nasal septum to extract primary chondrocytes that were then cultured and amplified. The passage 3 chondrocytes were used to construct ell sheets. Monolayer cell sheet was formed by intensive culturing and allowing the extracellular matrix secretion. Bilayer cell sheet was constructed by seeding passage 2 chondrocytes on the monolayer cell sheet. The cell sheets were harvested using cell scraper, their properties were investigated prior to plantation into nude mice to construct the tissue-engineered cartilage. RESULTS AND CONCLUSION: Both bilayer and monolayer cell sheets with soft tremellose structures showed no significant difference through naked eyes. The newly harvested cell sheets appeared to have good fluidity and gelation. Eight weeks after mplantation into the nude mice, mature cartilage blocks were obtained. Histologically, the cell sheets were thin films composed by layered chondrocytes and extracellular matrix. Glycosaminoglycan formation and type Ⅱ collagen expressions were observed in the cell sheets cultured in vitro. The explanted samples exhibited ature cartilaginous tissue at 8 weeks after implantation. Biochemical analysis showed that the DNA contents of the neocartilages were higher than those of native human costal cartilage, while the contents of glycosaminoglycan and hydroxyproline were similar to native human nasal septal cartilage. To conclude, the hondrocyte cell sheets are likely to be constructed and harvested successfully using common culture dish, and the cell sheets exhibit favourable chondrogenesis.关键词
组织构建/软骨组织工程/细胞膜片/软骨/培养/普通培养皿/糖胺聚糖/Ⅱ型胶原/成软骨性能/国家自然科学基金分类
医药卫生引用本文复制引用
周丽斌,吴玮,丁瑞英,徐冰心,韩浩伦,王刚,李保卫,王鸿南..软骨细胞膜片的成软骨性能[J].中国组织工程研究,2017,21(12):1843-1848,6.基金项目
国家自然科学基金青年科学基金项目(81401609)the National Natural Science Foundation of China, No. 81401609 （81401609）
