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Background : Genetic manipulation is an effective strategy to protect cells against environmental damages and enhance their capabilities for therapeutic usage. In order to avoid unwanted side effects, such as cancers, the expression of genes should be temporary increased. The aim of this study was to clone and temporary increased expression of a cell protective gene, Metallothionein 1 (MT1) in mesenchymal stem cells (MSCs) using plasmid expression system with the pcDNA 3.1 vector. Materials and Methods: Human bone marrow mesenchymal stem cells were isolated and validated by flow cytometry method. Complete cDNA sequence of MT1 gene was isolated and cloned in a plasmid vector named pcDNA 3.1. Recombinant gene construct was generated and transferred to the human MSCs. MT1 expression was monitored by RT-PCR and western blotting. Results: Recombinant human MT1 gene was successfully cloned in pcDNA vector and the correct format gene in the vector was confirmed by DNA sequencing. By RT-PCR and western blot methods, increased MT1 expression in MSCs was approved. The results showed that the expression of MT1 in the MSCs was transient. Conclusion: Genetic manipulation of MSCs by MT1using pcDNA 3.1 plasmid vector may be one of the protective strategies for transplanted cells from apoptosis and promote a new approach to provide an effective cell therapy after transplantation.

Experimental studies have shown that dental pulp tissue has potential to regenerate dentine in response to adverse stimuli, such as caries and associated operative procedures. However, the potential of dental pulp regeneration seems to be limited by regenerative capacity of the cell involved. In this study, we report the effect of transfection of a recombinant plasmid containing human BMP-2 gene in proliferation and differentiation of dental pulp tissue in vitro. The regenerative capacity was analyzed by ALP production and calcium content. Results showed that the transfected dental pulp cell was able to differentiate into the odontoblast phenotype, indicating the presence of odontoblast progentitor cells in dental pulp tissue.