Alireza Milani, Nooshin Haghighipour, Amin Hadi, Fatemeh Asgari, Azam Bolhassani: Enhancing the transfection potency of mesenchymal stem cells using heat shock method. The 3rd International and 11th National Biotechnology Congress, Tehran-Iran; 09/2019. (Poster)
Abstract Infection with specific subtypes of human papillomavirus (HPV) has been strongly implicated in cervical carcinogenesis. It appears that papillomavirus types 16 and 18 are associated with a higher risk of advanced cervical. HPV-E7 oncoprotein is necessary for development of cervical cancer and as a specific marker. Moreover, mesenchymal stem cells (MSCs) are unique bone marrow-derived progenitor cells that were recently used in therapeutic vaccines against cancer or infection diseases. The studies indicated that the efficiency of gene delivery in MSCs is very low; thus, some combinatorial methods such as chemical and physical approaches have attracted a special interest. In this study, MSCs were extracted from the long bone marrow of mice and identified by CD45 and CD90.2 surface markers using flow cytometry analysis. Then, the cells were transfected with pEGFP-E7/ lipofectamine complexes associated with heating at 42 °C for 2h and then incubated for 48 h at 37 °C and 5% CO2. Moreover, the transfection of MSCs with the plasmid was performed using lipofectamine alone. The flow cytometry results showed that the expression ratio of E7 gene transfected into MSCs using lipofectamine followed by heating was higher than lipofection alone. Indeed, heat shock led to enhance the penetration of the plasmid and further expression of E7 protein. Therefore, regarding the ability of modified MSC to express and secrete a viral antigen, the modified MSCs can be served to develop cell-based vaccines in future.
Keywords: HPV, Mesenchymal stem cells, In vitro transfection, Lipofectamine, Heat shock
