Three-Dimensional Vascular Network Assembly From Diabetic Patient-Derived Induced Pluripotent Stem Cells

Abstract

Objective In diabetics, hyperglycemia results in deficient endothelial progenitors and cells, leading to cardiovascular complications. We aim to engineer three-dimensional (3D) vascular networks in synthetic hydrogels from type-1 diabetes (T1D) patient-derived human induced pluripotent stem cells (hiPSCs), to serve as a transformative autologous vascular therapy for diabetic patients. Approach and Results We validated and optimized an adherent, feeder free differentiation procedure to derive early vascular cells (EVCs) with high portions of VEcad+ cells from hiPSCs. We demonstrate similar differentiation efficiency from hiPSCs derived from healthy donor and T1D patients. T1D-hiPSC-derived VEcad+ cells can mature to functional endothelial cells (ECs) expressing mature markers: von Willebrand factor and eNOS, are capable of lectin binding and acetylated low density lipoprotein uptake, form cords in Matrigel and respond to tumor necrosis factor alpha. When embedded in engineered hyaluronic acid (HA) hydrogels, T1D-EVCs undergo morphogenesis and assemble into 3D networks. When encapsulated in a novel hypoxia-inducible (HI) hydrogel, T1D-EVCs respond to low oxygen and form 3D networks. As xenografts, T1D-EVCs incorporate into developing zebrafish vasculature. Conclusion Using our robust protocol, we can direct efficient differentiation of T1D-hiPSC to EVCs. Early ECs derived from T1D-hiPSC are functional when mature. T1D-EVCs self-assembled into 3D networks when embedded in HA and HI hydrogels. The capability of T1D-EVCs to assemble into 3D networks in engineered matrices and to respond to a hypoxic microenvironment is a significant advancement for autologous vascular therapy in diabetic patients and has broad importance for tissue engineering.

Vascular Regeneration by Stem/Progenitor Cells

“…These endothelial cells could undergo morphogenesis and assemble into 3D networks when cultured in engineered hyaluronic acid hydrogels or in response to low oxygen in vivo and could incorporate into developing vasculature in zebrafish. 76 Similar results were also obtained in another study where human iPS cells derived from somatic tissue could differentiate into both endothelial cells and pericytes which were fully functional both in vivo and in vitro. 77 In investigation into the mechanisms of iPS cell differentiation, Chen et al 78 found that miR-199b is involved in endothelial differentiation.…”

“…75 Human iPS cells derived from patients with type 1 diabetes mellitus display a similar differentiation efficiency when compared with iPS cells derived from healthy donors. 76 These patient-derived iPS cells can differentiate into endothelial lineages expressing typical endothelial markers and with functional capabilities of lectin binding, acetylated low-density lipoprotein uptake, tube formation in matrigel, and response to TNF-α. These endothelial cells could undergo morphogenesis and assemble into 3D networks when cultured in engineered hyaluronic acid hydrogels or in response to low oxygen in vivo and could incorporate into developing vasculature in zebrafish.…”

Impact of Diabetes Mellitus

“…Chan and colleagues 17 were recently able to generate early vascular cells with high portions of vascular endothelial cadherin-positive cells from inducible pluripotent stem cells from patients with type 1 diabetes. These cells formed three-dimensional vascular networks in vitro and incorporated into the vasculature in a zebra fish model.…”

O2-controllable hydrogels for studying cellular responses to hypoxic gradients in three dimensions in vitro and in vivo

“…Those tissues with lower relative O 2 levels will exhibit shallower gradients 38 . We have demonstrated that insights into the role of varying O 2 during tumor growth, vascular development 42 , and angiogenesis 38,43 can be acquired using these hydrogels.…”

“…Thus, Gel-HI hydrogels provide a system that more closely mimics the native regenerative niche than do other culture systems. For example, our laboratory has shown that early vascular cells derived from induced pluripotent stem cells of diabetic patients can generate lumenized networks in hypoxic Gel-HI hydrogels, indicating their responsiveness to hypoxic pathways 42 .…”