OSTEOGENESIS OF GFP-TRANSFECTED HUMAN EMBRYONIC STEM CELLS

Abstract

Osteogenesis is one of the key components of bone development as well as regeneration. The use of human embryonic stem cells (hESCs) and fluorescent protein transfection has been respectively increased in the study of osteogenesis. However, the influence of fluorescent protein transfection on hESC osteogenesis still remains unknown. In this study, enhanced Green Fluoroscent Protein (eGFP) was transfected into H1 and H9 hESCs lines in a non-viral approach. The eGFP-transfected and non-transfected H1 and H9 were cultured in osteogenic media for 21 days. The osteogenesis of H1, H1eGFP+, H9 and H9eGFP+ were evaluated and compared by histochemical staining, immunocytochemical staining and real-time polymerase chain reaction. The expression of osteogenesis markers and osteoblast-specific markers indicated that the osteogenesis of H1eGFP+ and H9eGFP+ were not affected by eGFP transfection. The results have demonstrated the feasibility of lineage tracking via tagging hESCs with eGFP in the hESCs osteogenesis research.