Human induced pluripotent stem cells derived endothelial cells mimicking vascular inflammatory response under flow

Abstract

Endothelial cells (ECs) have great potential in vascular diseases research and regenerative medicine. Autologous human ECs are difficult to acquire in sufficient numbers in vitro, and human induced pluripotent stem cells (iPSCs) offer unique opportunity to generate ECs for these purposes. In this work, we present a new and efficient method to simply differentiate human iPSCs into functional ECs, which can respond to physiological level of flow and inflammatory stimulation on a fabricated microdevice. The endothelial-like cells were differentiated from human iPSCs within only one week, according to the inducing development principle. The expression of endothelial progenitor and endothelial marker genes (GATA2, RUNX1, CD34, and CD31) increased on the second and fourth days after the initial inducing process. The differentiated ECs exhibited strong expression of cellsspecific markers (CD31 and von Willebrand factor antibody), similar to that present in human umbilical vein endothelial cells. In addition, the hiPSC derived ECs were able to form tubular structure and respond to vascular-like flow generated on a microdevice. Furthermore, the human induced pluripotent stem cell-endothelial cells (hiPSC-ECs) pretreated with tumor necrosis factor (TNF-a) were susceptible to adhesion to human monocyte line U937 under flow condition, indicating the feasibility of this hiPSCs derived microsystem for mimicking the inflammatory response of endothelial cells under physiological and pathological process. V C 2016 AIP Publishing LLC. [http://dx

Determination of critical shear stress for maturation of human pluripotent stem cell‐derived endothelial cells towards an arterial subtype

“…Human pluripotent stem cell‐derived endothelial cells (hPSC‐ECs) on the other hand, are scalable and can be more readily expanded in vitro (Zhang et al, ). However, current differentiation protocols for hPSC‐ECs typically sort the differentiated population using only pan‐endothelial markers, such as CD‐31 (Levenberg, Golub, Amit, Itskovitz‐Eldor, & Langer, ; Rufaihah et al, ; Sivarapatna et al, ; L. Wang et al, ) or VE‐Cadherin (Adams et al, ; Liu et al, ), resulting in a highly heterogeneous population of cells that express arterial, venous, and lymphatic markers (Zhang et al, ). Such variability in EC phenotypes and functions inevitably limit the efficacy of their performance in endothelialized graft applications (Kudo et al, ; Rufaihah et al, ).…”

Induced Pluripotent Stem Cell–Derived Endothelial Cells

“…27 Methods to differentiate iPSC-ECs rely mostly on four approaches: co-culture, embryoid body formation, twodimensional culture with growth factors, and threedimensional (3D) culture (Table 1). 11,12,14,28 Earlier efforts to differentiate iPSCs into endothelial cells involved coculture with stromal cells. Although the local factors from bone marrow stromal cells resulted in differentiation into endothelial cells, the yield was low, with high variability of endothelial differentiation.…”

“…11 Although improved from the co-culture method, this embryoid body formation method yielded only 6% to 16% endothelial cells. 11 Wang et al 14 further expanded on the differentiation method via two-dimensional Matrigel culture. In the first differentiation stage, a combination of BMP4, activin A, basic fibroblast growth factor-2, and VEGF was used to drive the differentiation of mesoderm cells into cardiovascular cells.…”

“…For example, when iPSC-ECs were exposed to proinflammatory stimuli, such as IL-1b, TNF-a, and lipopolysaccharide, they expressed intercellular adhesion molecule 1, vascular cell adhesion molecule 1, and E-selectin. 11,14 Secretion of proinflammatory cytokines, such as monocyte chemotactic protein-1, IL-8, regulated on activation normal T cell expressed and secreted, and interferon-geinduced protein 10, was seen. 11 A barrier study in iPSC-ECs showed that histamine induced a transient increase in permeability and that VEGF induced a sustained increase in permeability in a monolayer of iPSC-ECs.…”

iPSC-derived endothelial cell response to hypoxia via SDF1a/CXCR4 axis facilitates incorporation to revascularize ischemic retina

“…Recent advances in induced pluripotent stem cell (iPSC) technology has allowed for robust procurement of vascular derivatives from stem cells sources. These iPSC-ECs offer a unique opportunity as an inexhaustible, constant source from which to generate cells for vascular therapies (7).…”