Genetically Engineered Mesenchymal Stem Cells Stably Expressing

Abstract

Objective(s)Mesenchymal stem cells (MSCs) are nonhematopoietic stromal cells that are capable of differentiating into and contribute to the regeneration of mesenchymal tissues. Human mesenchymal stem cells (hMSCs) are ideal targets in cell transplantation and tissue engineering. Enhanced green fluorescent protein (EGFP) has been an important reporter gene for gene therapy. The aim of this study was establishment of MSCs expressing GFP. Materials and MethodsMSCs were isolated and characterized by Immunophenotyping. The pEGFP-N1 plasmid was extracted from previously transformed Escherichia. coli cells and transfected into MSCs using FuGENE HD transfection reagent. Stable cells were established in the presence of geneticin. Expression of GFP was detected by RT-PCR, western blot analysis and immunoflorecent microscope. ResultsMSCs were successfully isolated and characterized. The MSCs transfected with the pEGFP-N1 plasmid expressed GFP both in mRNA and protein levels while cells transfected with empty vector did not. ConclusionThe results suggested that this engineered cell line will be used in the future studies and can easily be traced in vivo.

Keywords

• Genetic Engineering
• Green Fluorescent Protein
• Mesenchymal Stem Cells
• Transfection