Prospective isolation of human embryonic stem cell-derived cardiovascular progenitors that integrate into human fetal heart tissue

Abstract

A goal of regenerative medicine is to identify cardiovascular progenitors from human ES cells (hESCs) that can functionally integrate into the human heart. Previous studies to evaluate the developmental potential of candidate hESC-derived progenitors have delivered these cells into murine and porcine cardiac tissue, with inconclusive evidence regarding the capacity of these human cells to physiologically engraft in xenotransplantation assays. Further, the potential of hESC-derived cardiovascular lineage cells to functionally couple to human myocardium remains untested and unknown. Here, we have prospectively identified a population of hESC-derived ROR2 + / CD13 + /KDR + /PDGFRα + cells that give rise to cardiomyocytes, endothelial cells, and vascular smooth muscle cells in vitro at a clonal level. We observed rare clusters of ROR2 + cells and diffuse expression of KDR and PDGFRα in first-trimester human fetal hearts. We then developed an in vivo transplantation model by transplanting second-trimester human fetal heart tissues s.c. into the ear pinna of a SCID mouse. ROR2 + /CD13 + /KDR + /PDGFRα + cells were delivered into these functioning fetal heart tissues: in contrast to traditional murine heart models for cell transplantation, we show structural and functional integration of hESC-derived cardiovascular progenitors into human heart. engraftment | surface markers | Stem cells | mature cardiomyocytes | clonal analysis

How One Thing Led to Another

“…We thought if we generated a mouse whose granulocytes do not die on time and crossed it to CF mice, we might get clues as to how to ameliorate the disease and perhaps how to treat it with HSC transplantation from healthy donors. We cloned the upstream region of mrp8 and used it to drive human Bcl2 in transgenic mice (209 (212,213), but also triggered a variety of studies in the lab (152,201,(214)(215)(216)(217)(218)(219)(220)(221)(222)(223). For example, we demonstrated a number of developmental events wherein cytokine instructions acted by keeping a cell type alive to follow its intrinsic developmental progression, and the signal transduction pathway, e.g., monocytes to macrophages and osteoclasts in M-CSF-deficient osteopetrotic mice, could be replaced by stage-specific expression of bcl2 (52,214).…”

The effects of transplanted cells in stem cell therapy for myocardial ischemia

“…In clinical trials, it was shown that transplant of myoblasts could led to arrhythmia occurrence (Leobon et al, 2003). Additionally, a number of other studies also confirmed that transplanted beating heart cells could give rise to arrhythmia (Gillum and Sarvazyan, 2008;Menasche et al, 2008), while transplantation of human ESC (hESC)-derived cardiovascular progenitors into human fetal hearts showed harmony of the structure and function between transplanted cells and cardiac host cells (Ardehali et al, 2013). Moreover, the electromechanical coupling of hESC-derived cardiomyocytes and the suppression of cardiac arrhythmias in transplanted pig models seems to be related to the formation of electrical conduction bridges through scar tissues (Shiba et al, 2012).…”

We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells

“…Cells committed to the cardiac mesoderm lineage may be conveniently identified using only four surface markers -ROR2 (receptor tyrosine kinase-like orphan receptor 2, an inhibitor of Wnt canonical signalling); CD13 (alanine aminopeptidase, an antigen commonly present on the surface of leukocytes); VEGFR (KDR) and platelet-derived growth factor receptor alpha (PDGFRα, a cell-surface receptor for PDGF -A, B, and C) [178][179][180][181][182]. Human cells that are advanced into cardiogenic pathway of differentiation express cardio-specific proteins (mainly transcription factors) such as MESP1 (one of the first markers indicative of cardiogenesis), TBX5, GATA4, MEF2C, NKX2.5, and ISL1 [182].…”

“…Human cells that are advanced into cardiogenic pathway of differentiation express cardio-specific proteins (mainly transcription factors) such as MESP1 (one of the first markers indicative of cardiogenesis), TBX5, GATA4, MEF2C, NKX2.5, and ISL1 [182]. The expression of key cardiac transcription factors such as NKX2.5 and TBX5 ought to be detectable in the majority of the cells by the 4th day of differentiation [27].…”