Flk1+ and VE-Cadherin+ Endothelial Cells Derived from iPSCs Recapitulates Vascular Development during Differentiation and Display Similar Angiogenic Potential as ESC-Derived Cells

Abstract

RationaleInduced pluripotent stem (iPS) cells have emerged as a source of potentially unlimited supply of autologous endothelial cells (ECs) for vascularization. However, the regenerative function of these cells relative to adult ECs and ECs derived from embryonic stem (ES) cells is unknown. The objective was to define the differentiation characteristics and vascularization potential of Fetal liver kinase (Flk)1+ and Vascular Endothelial (VE)-cadherin+ ECs derived identically from mouse (m)ES and miPS cells. Methods and ResultsNaive mES and miPS cells cultured in type IV collagen (IV Col) in defined media for 5 days induced the formation of adherent cell populations, which demonstrated similar expression of Flk1 and VE-cadherin and the emergence of EC progenies. FACS purification resulted in 100% Flk1+ VE-cadherin+ cells from both mES and miPS cells. Emergence of Flk1+VE-cadherin+ cells entailed expression of the vascular developmental transcription factor Er71, which bound identically to Flk1, VE-cadherin, and CD31 promoters in both populations. Immunostaining with anti-VE-cadherin and anti-CD31 antibodies and microscopy demonstrated the endothelial nature of these cells. Each cell population (unlike mature ECs) organized into well-developed vascular structures in vitro and incorporated into CD31+ neovessels in matrigel plugs implanted in nude mice in vivo.ConclusionThus, iPS cell-derived Flk1+VE-cadherin+ cells expressing the Er71 are as angiogenic as mES cell-derived cells and incorporate into CD31+ neovessels. Their vessel forming capacity highlights the potential of autologous iPS cells-derived EC progeny for therapeutic angiogenesis.

EngineeringDe NovoAssembly of Fetal Pulmonary Organoids

“…In this context, fetal-pulmonary-cell-based systems will continue to be beneficial for building an understanding of the microenvironmental cues required to drive alveolar morphogenesis in vitro, while the stem cell field continues to work toward the derivation of the multiple epithelial, mesenchymal, and endothelial cell lineages required to build the distal lung. 12,13,75,76 In conclusion, we believe that harnessing crosstalk between endogenous networks of growth factor signals, such as the FGF/SHH/VEGF-A network, as well as focal patterns of synthesis and deposition of specific ECM molecules, such as TN-C, in concert with advances in scaffolding technologies and further progress in directed pulmonary differentiation of multifarious progenitor cells, will be essential stepping stones on the way to achieving the interdisciplinary goal of engineering complex functional human lung tissue for therapeutic and research purposes.…”

Successful re-endothelialization of a perfusable biological vascularized matrix (BioVaM) for the generation of 3D artificial cardiac tissue

“…Repopulation of the BioVaM vessel bed with human cord blood derived endothelial cells has been previously demonstrated [4]. The ideal solution would be the re-endothelialization with hiPSC-derived endothelial cells [1,17,31].…”

Induced Pluripotent Stem (iPS) Cell Culture Methods and Induction of Differentiation into Endothelial Cells

“…Therefore, in our laboratory, we have used iPS cells as a source for VE-cadherin+ and Flk1+ endothelial cells (ECs) and showed their ability to incorporate into CD31+ neovessels in Matrigel plugs (28) and into newly formed blood vessels in a mouse model of hind limb ischemia (28) and in Matrigel plug assays. Thus, based purely upon our recent publication (28), here, we outline methods for iPS culture, including the conditions used to differentiate iPS cells into ECs as well as for the isolation, purification, and characterization of VE-cadherin+ and Flk1+ ECs.…”