O-39: Expression of Cardiomyocyte Markers in Mouse Embryonic Fibroblasts by Cell Free Cardimyocyte Extract and Epigenetic Manipulation


Heidari F *, Talaei T , Rohani L , Esmaeil pour T , Vojdani Z , Mostafavipour Z ,

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Objective: Various cell populations, such as embryonic stem cells, cord blood cells, and mesenchymal stem cells, have been suggested as a source for cell therapy in curing heart diseases. Transdifferentiation of fully differentiated cells can be use as a potential source of cell therapy. In this study, we showed that heart extract could induce transdifferentiation of mouse embryonic fibroblasts (MEFs) into cardiomyocytes. Materials and Methods: MEFs were used as primary source of cells. Cardiomyocyte extract was prepared from adult mouse cardiomyocytes. MEFs were treated with 5-deoxyazacytidin and Trichostatin A. The treated cells were permeabilized with streptolysin O, and exposed to the mouse cardiomyocyte extract. The cells were cultured for 24 hours, 10 days and 21 days. Immunocytochemistry technique was performed for α-actinin, cardiac troponin T(CTT),Myosin Heavy Chain (MHC) and Atrial Nateriuretic Peptide (ANP) antibodies as cardiomyocyte markers. The results were compared with control group. Results: The results of immunocytochemistry for α-actinin, CTT, MHC and ANP antibodies revealed that After 24 hours showed no expression of this antibodies in all groups. After 10 days 75% of the cells were treated with 5-deoxyazacytidin and trichostatin A were expressed α-actinin and MHC.The results of immunocytochemistry for CTT and ANP antibodies showed no expression of this antibodies in all groups after 10 days. the results of immunocytochemistry for these antibodies show the expression of this markers in groups that treated with 5-deoxyazacytidin and Trichostatin A and cardiomyocyte extract after 21 days. Conclusion: Mouse embryonic fibroblasts expressed α-actinin and MHC, so that, it seems that the cardiomyocyte extract can induce tarnsdifferentiation of MEFs into cardiomyocyte. No expression of CTT and ANP after 24 hours and 10 days may arise of deficient maturity in reprogrammed MEFs. expression of α-actinin, MHC, CTT and ANP after 21 days show the time dependent expression of cardiac markers in somatic cells. This study shows that MEFs have the capability to use in the future research as a source of cells for reprogramming procedures