Novel Bioreactor Platform for Scalable Cardiomyogenic Differentiation from Pluripotent Stem Cell-Derived Embryoid Bodies
Generation of cardiomyocytes from pluripotent stem cells (PSCs) is a common and valuable approach to produce large amount of cells for various applications, including assays and models for drug development, cell-based therapies, and tissue engineering. All these applications would benefit from a reliable bioreactor-based methodology to consistently generate homogenous PSC-derived embryoid bodies (EBs) at a large scale, which can further undergo cardiomyogenic differentiation. The goal of this chapter is to describe a scalable method to consistently generate large amount of homogeneous and synchronized EBs from PSCs. This method utilizes a slow-turning lateral vessel bioreactor to direct the EB formation and their subsequent cardiomyogenic lineage differentiation.
Keywords:Bioreactor Cardiomyocyte Embryoid body Pluripotent stem cells Slow turning lateral vessel
The research that supports these methodologies was funded by grants from the EU FP7 (“PartnErS” PIAP-GA-2008-218205; “AniStem,” PIAP-GA-2011286264; “EpiHealth,” HEALTH-2012-F2-278418; “EpiHealthNet,” PITN-GA-2012-317146, “STEMMAD,” PIAPP-GA-2012-324451) Research Center of Excellence 9878/2015/FEKUT project, the Mahidol University, the Thailand Research Fund (TRF), the Office of Higher Education Commission, Thailand (OHEC), and the Mahidol University (MRG 5680108).
- 1.Chong JJ, Yang X, Don CW, Minami E, Liu YW, Weyers JJ, Mahoney WM, Van Biber B, Cook SM, Palpant NJ, Gantz JA, Fugate JA, Muskheli V, Gough GM, Vogel KW, Astley CA, Hotchkiss CE, Baldessari A, Pabon L, Reinecke H, Gill EA, Nelson V, Kiem HP, Laflamme MA, Murry CE (2014) Human embryonic-stem-cell-derived cardiomyocytes regenerate non-human primate hearts. Nature 510(7504):273–277. doi: 10.1038/nature13233 CrossRefPubMedPubMedCentralGoogle Scholar
- 2.Masumoto H, Ikuno T, Takeda M, Fukushima H, Marui A, Katayama S, Shimizu T, Ikeda T, Okano T, Sakata R, Yamashita JK (2014) Human iPS cell-engineered cardiac tissue sheets with cardiomyocytes and vascular cells for cardiac regeneration. Sci Rep 4:6716. doi: 10.1038/srep06716 CrossRefPubMedPubMedCentralGoogle Scholar
- 9.Rungarunlert S, Klincumhom N, Bock I, Nemes C, Techakumphu M, Pirity MK, Dinnyes A (2011) Enhanced cardiac differentiation of mouse embryonic stem cells by use of the slow-turning, lateral vessel (STLV) bioreactor. Biotechnol Lett 33(8):1565–1573. doi: 10.1007/s10529-011-0614-8 CrossRefPubMedGoogle Scholar
- 10.Rungarunlert S, Klincumhom N, Tharasanit T, Techakumphu M, Pirity MK, Dinnyes A (2013) Slow turning lateral vessel bioreactor improves embryoid body formation and cardiogenic differentiation of mouse embryonic stem cells. Cell Reprogram 15(5):443–458. doi: 10.1089/cell.2012.0082 CrossRefPubMedPubMedCentralGoogle Scholar
- 13.Muenthaisong S, Ujhelly O, Polgar Z, Varga E, Ivics Z, Pirity MK, Dinnyes A (2012) Generation of mouse induced pluripotent stem cells from different genetic backgrounds using Sleeping beauty transposon mediated gene transfer. Exp Cell Res 318(19):2482–2489. doi: 10.1016/j.yexcr.2012.07.014 CrossRefPubMedGoogle Scholar
- 14.Mauritz C, Schwanke K, Reppel M, Neef S, Katsirntaki K, Maier LS, Nguemo F, Menke S, Haustein M, Hescheler J, Hasenfuss G, Martin U (2008) Generation of functional murine cardiac myocytes from induced pluripotent stem cells. Circulation 118(5):507–517. doi: 10.1161/CIRCULATIONAHA.108.778795 CrossRefPubMedGoogle Scholar