In Vivo Generation of Post-infarct Human Cardiac Muscle by Laminin-Promoted Cardiovascular Progenitors

• Published in: Cell reports (Cambridge) 2019-03, Vol.26 (12), p.3231-3245.e9
• Main Authors: Yap, Lynn, Wang, Jiong-Wei, Moreno-Moral, Aida, Chong, Li Yen, Sun, Yi, Harmston, Nathan, Wang, Xiaoyuan, Chong, Suet Yen, Vanezis, Konstantinos, Öhman, Miina K., Wei, Heming, Bunte, Ralph, Gosh, Sujoy, Cook, Stuart, Hovatta, Outi, de Kleijn, Dominique P.V., Petretto, Enrico, Tryggvason, Karl
• Format: Article
• Language: English
• Subjects: Animals; cardiovascular progenitors; Disease Models, Animal; hESC-based cardiac muscle regeneration; Heterografts; Humans; laminin; Laminin - metabolism; Male; Medicin och hälsovetenskap; Mice; Mice, Nude; Myocardial Infarction - metabolism; Myocardial Infarction - pathology; Myocardial Infarction - therapy; Myocardium - metabolism; Myocardium - pathology; Myocytes, Cardiac - metabolism; Myocytes, Cardiac - pathology; Pluripotent Stem Cells - pathology; Pluripotent Stem Cells - transplantation; regenerative cardiology; Stem Cell Transplantation
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2019.02.083

Regeneration of injured human heart muscle is limited and an unmet clinical need. There are no methods for the reproducible generation of clinical-quality stem cell-derived cardiovascular progenitors (CVPs). We identified laminin-221 (LN-221) as the most likely expressed cardiac laminin. We produced it as human recombinant protein and showed that LN-221 promotes differentiation of pluripotent human embryonic stem cells (hESCs) toward cardiomyocyte lineage and downregulates pluripotency and teratoma-associated genes. We developed a chemically defined, xeno-free laminin-based differentiation protocol to generate CVPs. We show high reproducibility of the differentiation protocol using time-course bulk RNA sequencing developed from different hESC lines. Single-cell RNA sequencing of CVPs derived from hESC lines supported reproducibility and identified three main progenitor subpopulations. These CVPs were transplanted into myocardial infarction mice, where heart function was measured by echocardiogram and human heart muscle bundle formation was identified histologically. This method may provide clinical-quality cells for use in regenerative cardiology. [Display omitted] •LN-221 enables production of hESC-derived human cardiovascular progenitors•Bulk and single-cell RNA-seq data reveal highly reproducible differentiation•Cardiovascular progenitors survive in vivo and form human muscle fiber bundles•Improvement in heart function after cell transplantation Yap et al. perform in vitro differentiation of pluripotent embryonic stem cells to cardiovascular progenitors using laminin-221 as a culture matrix in a fully human and chemically defined protocol. Use of these cells for in vivo cardiac regeneration and to generate human heart muscle bundles results in improved heart function.