Granular Matrigel: restructuring a trusted extracellular matrix material for improved permeability

Abstract

Matrigel is a polymeric extracellular matrix material produced by mouse cancer cells. Over the past four decades, Matrigel has been shown to support a wide variety of two- and three-dimensional cell and tissue culture applications including organoids. Despite widespread use, transport of molecules, cells, and colloidal particles through Matrigel can be limited. These limitations restrict cell growth, viability, and function and limit Matrigel applications. A strategy to improve transport through a hydrogel without modifying the chemistry or composition of the gel is to physically restructure the material into microscopic microgels and then pack them together to form a porous material. These ‘granular’ hydrogels have been created using a variety of synthetic hydrogels, but granular hydrogels composed of Matrigel have not yet been reported. Here we present a drop-based microfluidics approach for structuring Matrigel into a three-dimensional, mesoporous material composed of packed Matrigel microgels, which we call granular Matrigel. We show that restructuring Matrigel in this manner enhances the transport of colloidal particles and human dendritic cells (DCs) through the gel while providing sufficient mechanical support for culture of human gastric organoids (HGOs) and co-culture of human DCs with HGOs.

Design and Development of Extracellular Matrix Protein‐Based Microcapsules as Tools for Bacteria Investigation

“…entactin, [34] the morphology of the obtained Matrigel-based Pro-Caps was different from the ProCaps consisting of laminin-111. Furthermore, Matrigel contains a broad mix of proteoglycans and growth factors.…”

“…First, Matrigel is widely used as a substrate for the growth and development of cells in 2D and 3D cultures. [ 34 ] Second, it is composed of laminin‐111 (60%), collagen type IV (30%), entactin (8%), and several growth factors such as TGF‐ β and IGF‐1. [ 34 ] Entactin serves as a bridging molecule between laminin‐111 and collagen type IV, and supports the structural organization of the Matrigel matrix.…”

“…[ 34 ] Second, it is composed of laminin‐111 (60%), collagen type IV (30%), entactin (8%), and several growth factors such as TGF‐ β and IGF‐1. [ 34 ] Entactin serves as a bridging molecule between laminin‐111 and collagen type IV, and supports the structural organization of the Matrigel matrix. [ 35 ] Using Matrigel allows us to investigate the establishment of protein microcapsules made out of a mixture of proteins and the interaction of organisms in a hollow confinement.…”

“…Following the analysis of E. coli behavior in ECM‐based ProCaps consisting of laminin‐111, we performed similar confocal time‐lapse experiments in the more complex Matrigel‐based ProCaps. It is important to mention here that due to a higher protein concentration and natural cross‐linkers such as entactin, [ 34 ] the morphology of the obtained Matrigel‐based ProCaps was different from the ProCaps consisting of laminin‐111. Furthermore, Matrigel contains a broad mix of proteoglycans and growth factors.…”

“…Furthermore, Matrigel contains a broad mix of proteoglycans and growth factors. [ 34 ] Matrigel capsules possess a more compact and rounded shape than the filamentous and elongated laminin‐111 ProCaps (Figure 2D).…”