Chondrogenesis of chondrocyte cell sheets

doi:10.3969/j.issn.2095-4344.2017.12.007

Abstract

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 culturing 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 cell 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 implantation 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 implantation 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 mature 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 chondrocyte cell sheets are likely to be constructed and harvested successfully using common culture dish, and the cell sheets exhibit favourable chondrogenesis.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Cartilage, Culture Techniques, Collage Type II, Tissue Engineering

CLC Number:

R318

Zhou Li-bin, Wu Wei, Ding Rui-ying, Xu Bing-xin, Han Hao-lun, Wang Gang, Li Bao-wei, Wang Hong-nan. Chondrogenesis of chondrocyte cell sheets[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(12): 1843-1848.

Figures/Tables 1

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