Versatile synthetic alternatives to Matrigel for vascular toxicity screening and stem cell expansion

Abstract

The physiological relevance of Matrigel as a cell-culture substrate and in angiogenesis assays is often called into question. Here, we describe an array-based method for the identification of synthetic hydrogels that promote the formation of robust in vitro vascular networks for the detection of putative vascular disruptors, and that support human embryonic stem cell expansion and pluripotency. We identified hydrogel substrates that promoted endothelial-network formation by primary human umbilical vein endothelial cells and by endothelial cells derived from human induced pluripotent stem cells, and used the hydrogels with endothelial networks to identify angiogenesis inhibitors. The synthetic hydrogels show superior sensitivity and reproducibility over Matrigel when evaluating known inhibitors, as well as in a blinded screen of a subset of 38 chemicals, selected according to predicted vascular disruption potential, from the Toxicity ForeCaster library of the US Environmental Protection Agency. The identified synthetic hydrogels should be suitable alternatives to Matrigel for common cell-culture applications.

Mechanical forces direct stem cell behaviour in development and regeneration

“…Recent work showed expansion of stem cells and development of structures in a synthetic PEG system with reversible crosslinks that are expected to impart viscoelasticity, although the mechanics were not yet characterized. 107 …”

Hydrogels in the clinic

“…Due to this high water content, hydrogels exhibit favorable biocompatibility and as such have been developed for and used in a variety of medical applications 1‐3 . Since hydrogel properties can be tuned through a variety of chemical approaches, their rational design and engineering has enabled new modalities for delivery of small molecules, 4‐7 proteins, 8‐12 and cells 13,14 and as tissue engineering scaffolds for directing cell fate/lineage, 15,16 stem cell expansion, 17‐20 and tissue regeneration 21‐25 . Research efforts to develop hydrogels for biomedical applications represents one of the most studied areas at the interface of engineering and medicine 26 .…”

Design and Fabrication of Three-Dimensional Printed Scaffolds for Cancer Precision Medicine

“…Nevertheless, Matrigel is generally not well defined and its application hinders detailed and accurate cell-extracellular matrix (ECM) interaction studies and creates issues with data reproducibility and translatability. 48 Moreover, Matrigel is a very soft material with inconsistent batch to batch variability and no direct control over the cell concentration and the physicochemical properties. 48 Therefore, investigation of ECM physical characteristics on cancer cell behaviors cannot be performed using Matrigel.…”

“…48 Moreover, Matrigel is a very soft material with inconsistent batch to batch variability and no direct control over the cell concentration and the physicochemical properties. 48 Therefore, investigation of ECM physical characteristics on cancer cell behaviors cannot be performed using Matrigel. Advances in biomaterials have enabled manufacturing natural and synthetic matrices with tunable biochemical, biophysical, and biomechanical properties in a range of physiological and pathophysiological tissues.…”

“…Advances in biomaterials have enabled manufacturing natural and synthetic matrices with tunable biochemical, biophysical, and biomechanical properties in a range of physiological and pathophysiological tissues. [48][49][50][51] These biomaterials can be optimized to support and promote organoid cultures with reproducible data. The control over the mechanical characteristics of biomaterials in organoids allows scientists to investigate and better understand the influence of matrix characteristics such as stuffiness on cells, which is challenging to be determined in vivo or using Matrigel-based organoids.…”