Selective Killing of Mouse Embryonic Stem Cells Using Reovirus
Date
2015-05-04
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Abstract
In the field of regenerative medicine, the use of pluripotent stem cells (PSC) to generate transplantable tissue is one of the leading methods undergoing investigation. The process of generating tissue derived from PSCs is not efficient and may leave pluripotent or progenitors cells behind following differentiation. These residual cells pose a tumorigenic threat when transplanted for therapeutic applications. In this study, we proposed a new approach utilizing reovirus to selectively target and kill PSCs. To study this, we utilized a murine embryonic stem cell and Type 3 Dearing reovirus system. We found mESCs treated with 16 multiplicity of infection (MOI) for a period of 2 days showed the greatest reduction in cell viability. In a mixture of mESCs and mouse embryonic fibroblasts (MEFs) we found that reovirus was selective in reducing the population of viable mESCs sparing MEFs. However, at high viral plaque forming units (PFU), reovirus has an inhibitory effect on MEFs as early as 3 days post infection. To elucidate the signalling pathway responsible for promoting reoviral killing of mESCs, we knocked out E-Ras, a homolog to Ras gene that is implicated in reoviral killing of cancer cells. We treated E-Ras KO mESCs with reovirus and found that E-Ras KO mESCs remain sensitive to reovirus treatment. Overall, we found that reovirus may not be an ideal method in selectively removing mESCs because of off target effects and also reovirus may be ineffective on hESCs.
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Medicine and Surgery, Engineering--Biomedical
Citation
Zhang, B. (2015). Selective Killing of Mouse Embryonic Stem Cells Using Reovirus (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/27744