Image-based crosstalk analysis of cell–cell interactions during sprouting angiogenesis using blood-vessel-on-a-chip

Abstract

Background Sprouting angiogenesis is an important mechanism for morphogenetic phenomena, including organ development, wound healing, and tissue regeneration. In regenerative medicine, therapeutic angiogenesis is a clinical solution for recovery from ischemic diseases. Mesenchymal stem cells (MSCs) have been clinically used given their pro-angiogenic effects. MSCs are reported to promote angiogenesis by differentiating into pericytes or other vascular cells or through cell–cell communication using multiple protein–protein interactions. However, how MSCs physically contact and move around ECs to keep the sprouting angiogenesis active remains unknown. Methods We proposed a novel framework of EC–MSC crosstalk analysis using human umbilical vein endothelial cells (HUVECs) and MSCs obtained from mice subcutaneous adipose tissue on a 3D in vitro model, microvessel-on-a-chip, which allows cell-to-tissue level study. The microvessels were fabricated and cultured for 10 days in a collagen matrix where MSCs were embedded. Results Immunofluorescence imaging using a confocal laser microscope showed that MSCs smoothed the surface of the microvessel and elongated the angiogenic sprouts by binding to the microvessel’s specific microstructures. Additionally, three-dimensional modeling of HUVEC–MSC intersections revealed that MSCs were selectively located around protrusions or roots of angiogenic sprouts, whose surface curvature was excessively low or high, respectively. Conclusions The combination of our microvessel-on-a-chip system for 3D co-culture and image-based crosstalk analysis demonstrated that MSCs are selectively localized to concave–convex surfaces on scaffold structures and that they are responsible for the activation and stabilization of capillary vessels.

Spatiotemporal observation reveals metastatic tumor-driven vascular remodeling as a potential route to polyclonal colonization

“…In-house polydimethylsiloxane-based chips (25 mm × 25 mm × 5 mm) were used to prepare the 3D MV model, as described previously [17,18]. A bovine serum albumin (BSA)-coated acupuncture needle (200 μm in diameter; Seirin, Shizuoka, Japan) was inserted into the chip to form a lumen structure in the type I collagen gel (2.4 mg/mL, Cellmatrix type I-A, Nitta Gelatin, Osaka, Japan).…”

The Edifice of Vasculature-On-Chips: A Focused Review on the Key Elements and Assembly of Angiogenesis Models

“…Collagen is arguably the most widely used hydrogel material for OoC angiogenesis models. − One primary factor is that the composition and microstructure of collagen-based hydrogels were found to help regulate vascular network formation . Other studies have identified that ADSCs can accelerate angiogenesis of ECs through matrix metalloproteinase (MMP)-mediated proteolytic collagen remodeling .…”

Advances in organ‐on‐a‐chip for the treatment of cardiovascular diseases

“…On this basis, the authors applied techniques such as 2D and 3D image analysis to investigate the specific mechanisms and spatiotemporal behaviors of MSCs during angiogenesis (Figure 6A). 122 In another example, Brouns and colleagues developed a novel vessel‐on‐a‐chip with a monolayer of ECs that could be used to investigate the antiplatelet and anticoagulation properties of ECs under whole blood flow conditions 123 . Recently, Wang developed a 3D atherosclerosis model using GelMA hydrogel, which is based on a multilayered vascular structure consisting of ECs and SMCs, with a hyperlipidemic environment and inflammatory stimulation as biochemical conditions for AS predisposition, thus providing a bionic platform to understand the progression of AS and the assessment of zinc ions released from zinc‐based scaffolds (Figure 6B).…”

“…(A) Conceptual sketch of this study to analyze angiogenic effects of MSC co cultured on the microvessel on a chip system. Reprinted with permission 122 . Copyright 2022, The Authors.…”