Abstract
Human pluripotent stem cell (hPSC)-derived cardiomyocytes (CMs) hold great promise for the repair of the injured heart, but optimal cell production in a fully chemically defined and cost-effective system is essential for the efficacy and safety of cell transplantation therapies. In this study, we provided a simple and efficient strategy for cardiac differentiation from hPSCs and performed functional evaluation in a rat model of myocardial infarction. Using a chemically defined medium including four components, recombinant human albumin, ascorbic acid, human transferrin, and RPMI 1640, we developed a manageable and cost-effective protocol for robust generation of CMs from hPSCs. Interestingly, the addition of transferrin helped hPSCs to transit from TeSR-E8 medium to the simple cardiac differentiation medium and successfully initiated mesoderm differentiation without significant cell death. The CM generation efficiency was up to 85% based on cTnT expression. We performed transcriptome profiling from differentiation day 0 to 35, and characterized interesting dynamic change of cardiac genes. CMs derived from transferrin-supplemented simple medium have similar transcriptome and the maturation level compared to those generated in B27 minus insulin medium as well as their in vivo counterparts. Importantly, after transplantation, hPSC-derived CMs survived in the infarcted rat heart, significantly improved the physiological function and reduced fibrosis. Our study offers an easy-to-use and cost-effective method for cardiac differentiation and facilitates the translational application of hPSC-derived CMs for heart repair.
Highlights
Adult mammalian hearts have a limited regenerative capacity, loss of cardiomyocytes is a leading cause of heart failure (Laflamme and Murry 2011; Weinberger and Eschenhagen 2020)
Both hESCs and human induced pluripotent stem cells (hiPSCs) maintained in TeSR-E8 medium were digested into single cells by Accutase (Millipore) and plated onto Matrigel-coated culture dishes at a density of 1–2 × 104 cells/cm2 in TeSR-E8 medium with 10 μM Y27632
All animal procedures in this study were approved by the Institutional Animal Care and Use Committee (IACUC) of the Chinese Academy of Military Medical Science
Summary
Adult mammalian hearts have a limited regenerative capacity, loss of cardiomyocytes is a leading cause of heart failure (Laflamme and Murry 2011; Weinberger and Eschenhagen 2020). B27 is a complex mix containing 21 components, many of which are derived from animal origin. Journal of Molecular Histology (2021) 52:87–99 animal-derived factors to human cardiomyocyte production, such as impeding elucidation of underlying mechanism, influencing reproducibility of differentiation, as well as increasing health risks for downstream application. We observed that the sudden transition from TeSR-E8 medium which contains high levels of insulin and transferrin to CDM3 frequently led to a marked increase of cell death, and reduced the final CM yield. We reasoned that the addition of the transferrin might be beneficial for cell survival and smooth transition from TeSR-E8 medium to simple cardiac differentiation medium like CDM3. The transferrin supplement enabled hPSCs to go through cardiac differentiation without much cell loss. Upon transplantation into a rat model of myocardial infarction (MI), hPSC-derived CMs improved remuscularization of the infarcted area and the function of the heart
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