Cardiomyocyte marker expression in a human lymphocyte cell line using mouse cardiomyocyte extract

Abstract

Cell transplantation shows potential for the treatment of cardiac diseases. Embryonic stem cells, cord blood and mesenchymal stem cells have been suggested as sources for transplantation therapy. Because of some technical limitations with the use of stem cells, transdifferentiation of fully differentiated cells is a potentially useful alternative. We investigated whether human peripheral blood cells could transdifferentiate into cardiomyocyte. Transdifferentiation was induced in a human B lymphocyte cell line (Raji). Cardiomyocyte extract was prepared from adult mouse cardiomyocytes. The cells were treated with 5-aza-2-deoxycytidine and trichostatin A, permeabilized with streptolysin O, and exposed to the mouse cardiomyocyte extract. They were cultured for 10 days, 3 weeks and 4 weeks. Cardiomyocyte markers were detected with immunohistochemistry and flow cytometry. Immunocytochemistry revealed that some cells expressed myosin heavy chain, α-actinin and cardiac troponin T after 3 and 4 weeks. Flow cytometry confirmed these data. In cells exposed to trichostatin A and 5-aza-2-deoxycytidine and permeabilized in the presence of the cardiomyocyte extract, troponin T expression was seen in 3.53% of the cells and 3.11% of them expressed α-actinin. After exposure to the cardiomyocyte extract, some permeabilized cells adhered to the plate loosely; however, the morphology did not change significantly, and they continued to show a rounded shape after 4 weeks. Our treated lymphocytes expressed cardiomyocyte markers. Our results suggest that lymphocytes may be useful in future research as a source of cells for reprogramming procedures.

Role of spermatogonial stem cells extract in transdifferentiation of 5-Aza-2′-deoxycytidine-treated bone marrow mesenchymal stem cells into germ-like cells

“…To explain this, it could be said that 5‐aza‐dC is more effective in MSC S differentiation into other cell types (Fukuda, ; Makino et al, ; Wakitani et al, ; Xu et al, ). It seems that in application of 5‐aza‐dC more time is needed to get an acceptable response (Taranger et al, ), and the better results may be obtained by using hyper‐acetylating agents in combination with 5‐aza‐dC (Talaei‐Khozani et al, ; Talaei‐Khozani et al, ; Vojdani et al, ); furthermore, 5‐aza‐dC would be more effective on ESCs markers expression, such as OCT4, SOX2 and Nanog (Hattori et al, ; Talaei‐Khozani et al, ; Tsuji‐Takayama et al, ), and its usage in most of the studies was on cell lines, unlike our study which was on primary cell cultures (Diepeveen et al, ; Håkelien et al, ). Additionally, 5‐aza‐dC at diverse doses exerts various effects on cell morphology and gene expression (Kumar et al, ; Zhou et al, ), and also epigenetic status is varied in dissimilar animal cells (Kang et al, ), and changed with time and with environmental stimulations (Jirtle and Skinner, ; Szyf, ).…”

The Effects of Chicken Embryo Brain Extract on Sciatic Nerve Regeneration of Male Rat; An Experimental Study

“…It has been shown that the extract from the differentiated cells change that fate of the other cell types [ 8 ]. In addition, it can promote cell reprogramming in somatic cells such as fibroblasts [ 9 ], lymphocytes [ 10 ], and granulosa cells [ 11 ]. Talaei-Khozani et al ., [ 12 ] showed that fibroblasts were also able to express cardiomyocyte markers by extract treatment.…”