c-Kit+ progenitors generate vascular cells for tissue-engineered grafts through modulation of the Wnt/Klf4 pathway

Abstract

The development of decellularised scaffolds for small diameter vascular grafts is hampered by their limited patency, due to the lack of luminal cell coverage by endothelial cells (EC) and to the low tone of the vessel due to absence of a contractile smooth muscle cells (SMC). In this study, we identify a population of vascular progenitor c-Kit+/Sca-1- cells available in large numbers and derived from immuno-privileged embryonic stem cells (ESCs). We also define an efficient and controlled differentiation protocol yielding fully to differentiated ECs and SMCs in sufficient numbers to allow the repopulation of a tissue engineered vascular graft. When seeded ex vivo on a decellularised vessel, c-Kit+/Sca-1-derived cells recapitulated the native vessel structure and upon in vivo implantation in the mouse, markedly reduced neointima formation and mortality, restoring functional vascularisation. We showed that Krüppel-like transcription factor 4 (Klf4) regulates the choice of differentiation pathway of these cells through β-catenin activation and was itself regulated by the canonical Wnt pathway activator lithium chloride. Our data show that ESC-derived c-Kit+/Sca-1-cells can be differentiated through a Klf4/β-catenin dependent pathway and are a suitable source of vascular progenitors for the creation of superior tissue-engineered vessels from decellularised scaffolds.

Follistatin-Like 3 Enhances the Function of Endothelial Cells Derived from Pluripotent Stem Cells by Facilitating β-Catenin Nuclear Translocation Through Inhibition of Glycogen Synthase Kinase-3β Activity

“…In this study, using the model of iPS‐ECs generation, FSTL3 expression was found to be upregulated by KLF4 , a key mediator of EC differentiation . In addition, FSTL3 expression progressively increased throughout the iPS‐ECs generation process, highlighting a likely angiogenic role.…”

“…A). We and others have previously shown that KLF4 is implicated in EC differentiation from pluripotent stem cells. In our iPS‐ECs system, it was confirmed that KLF4 was increased during EC differentiation, with a concomitant decrease of the pluripotent marker OCT4 (Fig.…”

Mesenchymal stem cells and vascular regeneration

“…When such engineered vessels were grafted in vivo , they demonstrated better patency than cell‐free scaffolds . A recent study utilized embryonic stem cell‐derived c‐kit+ cells as a source for SMCs and ECs and got encouraging results, with increased patency of in vivo grafted vessels for the generation of bioengineered vascular grafts . However, the application of embryonic stem cells is largely hampered by ethical concerns and induced pluripotent stem cells require the transfection of pluripotency factors often via viral delivery.…”

Human Peripheral Blood-Derived Endothelial Colony-Forming Cells Are Highly Similar to Mature Vascular Endothelial Cells yet Demonstrate a Transitional Transcriptomic Signature

“…Distinct endothelial lineages have considerable differences regarding their transcriptome, and the extent of endothelial cell heterogeneity is currently a matter of debate [17][18][19]. Nevertheless, endothelial identity is mandatory to control vascular tone and for the paracrine regulation of vascular specification within the graft [20][21][22]. Accordingly, transcriptomic signatures of pre-seeded endothelial cells (EC) should correspond to those of mature resident EC.…”