SUMMARY. Cell transplantation is a modern strategy for injured heart tissue regeneration. One of promising sources of cardiomyocytes is embryonic stem cells (ESCs) and induced pluripotent stem cells (IPSCs). Genetically modified murine ESC and IPSC expressing green fluorescent protein (GFP) under the control of a cardio specific α-MHC promoter were used for the research; differentiation was carried out in the suspension culture with constant stirring. The effect of dimethylsulfoxide (DMSO) on the processes of differentiation of pluripotent stem cells into cardiomyocytes was studied both at the stage of mesoderm cells formation and at the stage of the cardiomyocytes progenitors formation. Optimal term of DMSO application into the culture was determined. It was established that the addition of DMSO from the 5th to the 9th day of cultivation increased the efficiency of cardiomyocytes differentiation by one and a half times.
Keywords: embryonic stem cells, induced pluripotent stem cells, cardiomyocytes, dimethylsulfoxide, differentiation
Full text and supplemented materials
Free full text: PDFReferences
1. Soonpaa, M.H., Koh, G.Y., Klug, M.G., and Field, L.J., Formation of nascent intercalated disks between grafted fetal cardiomyocytes and host myocardium, Science, 1994, vol. 264, no. 5155, pp. 98–101. https://doi.org/10.1126/science.8140423
2. Leor, J., Patterson, M., Quinones, M.J., Kedes, L.H., and Kloner, R.A., Transplantation of fetal myocardial tissue into the infracted myocardium of rat. A potential method for repair of infarcted myocardium?, Circulation, 1996, vol. 94, no. 9, pp. 332–336.
3. Kehat, I., Kenyagin-Karsenti, D., Snir, M., Segev, H., Amit, M., Gepstein, A., Livne, E., Binah, O., Itskovitz-Eldor, J., and Gepstein, L., Human embryonic stem cells can differentiate into myocytes with structural and functional properties of cardiomyocytes, J. Clin. Invest., 2001, vol. 108, no. 3, pp. 407–414. https://doi.org/10.1172/JCI12131
4. Evans, M.J. and Kaufman, M.H., Establishment in culture of pluripotential cells from mouse embryos, Nature, 1981, vol. 292, pp. 154–156.
5. Takahashi, K. and Yamanaka, S., Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors, Cell, 2006, vol. 126, pp. 663–676. https://doi.org/10.1016/j.cell.2006.07.024
6. Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T., Tomoda, K., and Yamanaka, S., Induction of pluripotent stem cells from adult human fibroblasts by defined factors, Cell, 2007, vol. 131, no. 5, pp. 861–872.
7. Ebben, J.D., Zorniak, M., Clark, P.A., and Kuo, G.S., Introduction to induced pluripotent stem cells: advancing the potential for personalized medicine, World Neurosurg., 2011, vol. 76, nos. 3–4, pp. 270–275.
8. Vitalea, A.M., Wolvetangb, E., and Mackay-Sima, A., Induced pluripotent stem cells: A new technology to study human diseases, Int. J. Biochem. Cell Biol., 2011, vol. 43, pp. 843–846.
9. Jasmin Spray, D.C., Campos de Carvalho, A.C., and Mendez-Otero. R., Chemical induction of cardiac differentiation in P19 embryonal carcinoma stem cells, Stem. Cells Dev., 2010, vol. 19, no. 3, pp. 403–411. https://doi.org/10.1089/scd.2009.0234
10. Inamdar, M.S., Venu, P., Srinivas, M.S., Rao, K., and Vijay Raghavan, K., Derivation and characterization of two sibling human embryonic stem cell lines from discarded grade III embryos, Stem. Cells Dev., 2009, vol. 18, no. 3, pp. 423–433. https://doi.org/10.1089/scd.2008.0131
11. van der Heyden, M.A. and Defize, L.H., Twenty one years of P19 cells: what an embryonal carcinoma cell line taught us about cardiomyocyte differentiation, Cardiovasc Res., 2003, vol. 58, no. 2, pp. 292–302.
12. Santos, N.C., Figueira-Coelho, J., Martins-Silva, J., and Saldanha, C., Multidisciplinary utilization of dimethyl sulfoxide: pharmacological, cellular, and molecular aspects, Biochem. Pharmacol., 2003, vol. 65, no. 7, pp. 1035–1041.
13. Pal, R., Mamidi, M.K., Das, A.K., and Bhonde, R., Diverse effects of dimethyl sulfoxide (DMSO) on the differentiation potential of human embryonic stem cells, Arch. Toxicol., 2012, vol. 86, no. 4, pp. 651–661. https://doi.org/10.1007/s00204-011-0782-2
14. Meissner, A., Wernig, M., and Jaenisch, R., Direct reprogramming of genetically unmodified fibroblasts into pluripotent stem cells, Nat. Biotechnol., 2007, vol. 25, no. 10, pp. 1177–1181. https://doi.org/10.1038/nbt1335
15. Kolossov, E., Bostani, T., Roell, W., Breitbach, M., Pillekamp, F., Nygren, J.M., Sasse, P., Rubenchik, O., Fries, J.W., Wenzel, D., Geisen, C., Xia, Y., Lu, Z., Duan, Y., Kettenhofen, R., Jovinge, S., Bloch, W., Bohlen, H., Welz, A., Hescheler, J., Jacobsen, S.E., and Fleischmann, B.K., Engraftment of engineered ES cell-derived cardiomyocytes but not BM cells restores contractile function to the infracted myocardium, J. Exp. Med., 2006, vol. 203, pp. 2315–2327. https://doi.org/10.1084/jem.20061469
16. Fatima, A., Xu, G., Nguemo, F., Kuzmenkin, F., Burkert, K., Hescheler, J., and Šarić, T., Murine transgenic iPS cell line for monitoring and selection of cardiomyocytes, Lab Resource: Stem Cell Line, 2016, vol. 17, no. 2, pp. 266–272. https://doi.org/10.1016/j.scr.2016.07.007
17. Marks, P.A. and Breslow, R., Dimethyl sulfoxide to vorinostat: development of this histone deacetylase inhibitor as an anticancer drug, Nat. Biotechnol., 2007, vol. 25, no. 1, pp. 84–90. https://doi.org/10.1038/nbt1272
18. Johnstone, R.W., Histone-deacetylase inhibitors: novel drugs for the treatment of cancer, Nat. Rev. Drug. Discov., 2002, vol. 1, no. 4, pp. 287–299. https://doi.org/10.1038/nrd772
19. Chetty, S., Pagliuca, F.W., Honore, C., Kweudjeu, A., Rezania, A., and Melton, D.A., A simple tool to improve pluripotent stem cell differentiation, Nat. Methods, 2013, vol. 10, no. 6, pp. 553–556. https://doi.org/10.1038/nmeth.2442
20. Czysz, K., Minger, S., and Thomas, N., DMSO efficiently down regulates pluripotency genes in human embryonic stem cells during definitive endoderm derivation and increases the proficiency of hepatic differentiation, PLoS One, 2015, vol. 10, no. 2, pp. 1–16. https://doi.org/10.1371/journal.pone.0117689