Human Induced Pluripotent Stem Cell-Derived Endothelial Cells for Three-Dimensional Microphysiological Systems

Abstract

Microphysiological systems (MPS), or "organ-on-a-chip" platforms, aim to recapitulate in vivo physiology using small-scale in vitro tissue models of human physiology. While significant efforts have been made to create vascularized tissues, most reports utilize primary endothelial cells that hinder reproducibility. In this study, we report the use of human induced pluripotent stem cell-derived endothelial cells (iPS-ECs) in developing three-dimensional (3D) microvascular networks. We established a CDH5-mCherry reporter iPS cell line, which expresses the vascular endothelial (VE)-cadherin fused to mCherry. The iPS-ECs demonstrate physiological functions characteristic of primary endothelial cells in a series of in vitro assays, including permeability, response to shear stress, and the expression of endothelial markers (CD31, von Willibrand factor, and endothelial nitric oxide synthase). The iPS-ECs form stable, perfusable microvessels over the course of 14 days when cultured within 3D microfluidic devices. We also demonstrate that inhibition of TGF-β signaling improves vascular network formation by the iPS-ECs. We conclude that iPS-ECs can be a source of endothelial cells in MPS providing opportunities for human disease modeling and improving the reproducibility of 3D vascular networks.

Generation of reporter hESCs by targeting EGFP at the CD144 locus to facilitate the endothelial differentiation

“…Kurokawa et al. () constructed a CDH5‐mCherry iPSC line by generating CDH5 and mCherry fusion gene. The fusion protein may impact the permeability and stability.…”

“…EGFP is linked to the CDH5 gene by a 2A linker peptide, so the EGFP expression depended on the cleavage efficiency of 2A peptides. Different 2A peptides have different cleavage efficiencies and this may influence the gene expression behind the 2A peptides (Kim et al, 2011). Kurokawa et al (2017 constructed a CDH5-mCherry iPSC line by generating CDH5 and mCherry fusion gene.…”

Recapitulating the Vasculature Using Organ-On-Chip Technology

“…This process starts with angioblasts (endothelial precursor cells) differentiating at the edge of small blood islands ( Figure 2A1) [4,5,11]. These precursor cells form small clumps of differentiated cells (endothelial cells) and start invading the tissue with small extensions from the main mass ( Figure 2A2) [4,12]. These extensions of cells grow towards each other and form connections between the cell masses, forming a rudimentary network of endothelial cells ( Figure 2A3) [4,[8][9][10][11][12].…”

“…These precursor cells form small clumps of differentiated cells (endothelial cells) and start invading the tissue with small extensions from the main mass ( Figure 2A2) [4,12]. These extensions of cells grow towards each other and form connections between the cell masses, forming a rudimentary network of endothelial cells ( Figure 2A3) [4,[8][9][10][11][12]. This network opens up for perfusion and is further stabilised via maturation (see maturation, below) ( Figure 2A4) [4,5,8,[10][11][12].…”

Evaluating natural killer cell cytotoxicity against solid tumors using a microfluidic model

“…[29][30][31] Different microfluidic models have been used to recreate the tumor microenvironment and key processes including tumorinduced angiogenesis during cancer metastasis. [32][33][34][35] Recently, some models have been proposed to study the interaction between immune cells and solid tumors; focusing on the effect of hypoxia on immune migration or T cell receptor modification. [36][37][38] In this work, we present a microfluidic model to study NK cell immunotherapies and ADCC.…”