Abstract

Introduction: The osteoarthritis is a serious threat for well-developed and ageing countries. Present techniques of treatment of damaged cartilage are not sufficient. Hence, new strategies should be developed. One of the potential sources for the regeneration of cartilage is pluripotent stem cells (PSC). Aim: The development of an efficient protocol of chondrogenic differentiation using PSC. Material and methods: The human embryonic stem cell line (BG01V) was used in this study. The chondrogenic differentiation was performed using high-density pellet culture in the presence of TGF-β1 (10 ng/ml) and BMP2 (100 ng/ml). After 21 days gene expression analysis of markers related to chondrogenesis was done. Additionally, the histological staining was performed to detect the deposition of proteoglycans and collagens in differentiated pellet culture. Results: Obtained pellets exhibited decreased expression of pluripotent markers. The upregulation of mesodermal marker and type II collagen was observed in differentiated pellets in the presence of applied growth factors. The histological analysis revealed mild deposition of proteoglycans and collagens. Conclusion: The presented approach enables to obtain chondrogenic pellets in their early stages of chondrogenesis.

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