Hypoxia effects on the chondrogenic differentiation of three-dimensional co-cultured adipose-derived stem cells and articular chondrocytes

doi:10.3969/j.issn.2095-4344.2014.29.007

Abstract

Abstract:

BACKGROUND: Many in vivo and in vitro experiments indicate that hypoxic co-cultures promote stem cells differentiate into chondrocytes.

OBJECTIVE: To evaluate the influence of hypoxia on the chondrogenic differentiation of three-dimensional co-cultured adipose-derived stem cells and articular chondrocytes.

METHODS: Adipose-derived stem cells and articular chondrocytes were mixed at the ratio of 3:1, then the mixed cells were seeded onto poly(lactic-co-glycolic acid)-gelatin scaffold at the ultimate concentration of 5.0×10¹⁰/L. The cells were cultured in normoxia (20% O₂) and hypoxic (5% O₂) conditions for 6 weeks. After culture, hematoxylin and eosin staining was performed for histological structure analysis, and alcian blue staining was used to evaluate glycosaminoglycan synthesis. Type II collagen expression was detected by immunohistochemistry staining. The content of DNA, glycosaminoglycan and hydroxyproline in the scaffold-cell complex was measured.

RESULTS AND CONCLUSION: In the hypoxia group, hematoxylin-eosin staining showed the formation of massive cells and extracellular matrix; alcian blue staining showed massive glycosaminoglycan formation; immunohistochemistry staining detected strongly positive expression of collagen type II, the content of DNA, glycosaminoglycan and hydroxyproline was higher than the normoxia group. Hypoxia promotes in vitro chondrogenic differentiation of co-cultured adipose-derived stem cells and articular chondrocytes.

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

全文链接：

Key words: stem cells, adipose tissue, chondrocytes, tissue scaffolds, co-culture techniques, cell hypoxia

CLC Number:

R318

Dai Bing, Xu Hai-ting, Jin Hai-dong, Chen Hui, Cai Jian-wu, Fan Shi-yang, Pan Jun. Hypoxia effects on the chondrogenic differentiation of three-dimensional co-cultured adipose-derived stem cells and articular chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(29): 4630-4635.

Figures/Tables 2

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