Human iPSC-Derived Vascular Smooth Muscle Cells in a Fibronectin Functionalized Collagen Hydrogel Augment Endothelial Cell Morphogenesis

Abstract

Tissue-engineered constructs have immense potential as autologous grafts for wound healing. Despite the rapid advancement in fabrication technology, the major limitation is controlling angiogenesis within these constructs to form a vascular network. Here, we aimed to develop a 3D hydrogel that can regulate angiogenesis. We tested the effect of fibronectin and vascular smooth muscle cells derived from human induced pluripotent stem cells (hiPSC-VSMC) on the morphogenesis of endothelial cells. The results demonstrate that fibronectin increases the number of EC networks. However, hiPSC-VSMC in the hydrogel further substantiated the number and size of EC networks by vascular endothelial growth factor and basic fibroblast growth factor secretion. A mechanistic study shows that blocking αvβ3 integrin signaling between hiPSC-VSMC and fibronectin impacts the EC network formation via reduced cell viability and proangiogenic growth factor secretion. Collectively, this study set forth initial design criteria in developing an improved pre-vascularized construct.

Human iPSC‐Vascular smooth muscle cell spheroids demonstrate size‐dependent alterations in cellular viability and secretory function

“…To realize their full therapeutic potential, there remains a need to optimize culture conditions to improve cell survival, and paracrine secretion 28 . We and others have used functionalized biomaterials to enhance stem cell survival, and paracrine secretion both in vitro and in vivo 16,37–39 . Spheroids with a 3D tissue‐like architecture have been shown to improve cell‐survival, paracrine secretion, and improved regenerative potential 4,6,8,14,16,21,28 …”

“…A pure population of hiPSC‐VSMC was generated using a previously established protocol 1,37–39 . In brief, hiPSCs were cultured in feeder‐free conditions until reaching 80% confluency.…”

“…Our previous work has shown that the hiPSC‐VSMC secretome improves cell‐mediated regenerative healing 1,37–39 . We showed that their secretome improved wound closure and angiogenesis via secretion of proangiogenic growth factors in both acute and diabetic wound models 37–41 . Others have used hiPSC‐VSMC to develop structures resembling microvasculature to be used in tissue engineering 42,43 .…”

“…28,35,36 Our previous work has shown that the hiPSC-VSMC secretome improves cell-mediated regenerative healing. 1,[37][38][39] We showed that their secretome improved wound closure and angiogenesis via secretion of proangiogenic growth factors in both acute and diabetic wound models. [37][38][39][40][41] Others have used hiPSC-VSMC to develop structures resembling microvasculature to be used in tissue engineering.…”

“…1,[37][38][39] We showed that their secretome improved wound closure and angiogenesis via secretion of proangiogenic growth factors in both acute and diabetic wound models. [37][38][39][40][41] Others have used hiPSC-VSMC to develop structures resembling microvasculature to be used in tissue engineering. 42,43 The benefits of hiPSC-VSMC encourages us to explore mechanisms to modulate the paracrine secretion and in turn, their therapeutic potential in wound healing.…”