Micro‐Engineered 3D Scaffolds for Cell Culture Studies

Abstract

Cells in physiological 3D environments differ considerably in morphology and differentiation from those in 2D tissue culture. Naturally derived polymer systems are frequently used to study cells in 3D. These 3D matrices are complex with respect to their chemical composition, mechanical properties, and geometry. Therefore, there is a demand for well-defined 3D scaffolds to systematically investigate cell behavior in 3D. Here, fabrication techniques, materials, architectures, biochemical functionalizations, and mechanical properties of 3D scaffolds are discussed. In particular, work focusing on single cells and small cell assemblies grown in tailored synthetic 3D scaffolds fabricated by computer-based techniques are reviewed and the influence of these environments on cell behavior is evaluated.

Cell type-specific adaptation of cellular and nuclear volume in micro-engineered 3D environments

“…Simultaneous absorption of two photons leads to the excitation of photo-initiator molecules, which then trigger a highly localized chemical polymerization event that is confined to the focal volume of the laser due to the non-linearity of the 2PP process (Supplemental Fig S1) [12,28,40,42]. The spot size for the 2PP is diffraction limited and in our setup has a size of approx.…”

“…To overcome some of the limitations of artificial planar and rigid cell culture substrate, we [32][33][34][35][36] and others [12,28,[37][38][39][40] have used the direct laser writing (DLW) photopolymerization technique to fabricate tailored 3D scaffolds for single cell cultivation. By using polymers with different mechanical properties or by adjusting scaffold feature sizes, the stiffness of DLW-produced cellular micro-environments can be accurately adjusted [35].…”

Fabrication of 3D cell-laden hydrogel microstructures through photo-mold patterning

“…Each cell continuously interacts with its surrounding 3D microenvironment through biochemical, biomechanical and bioelectrical signals, which vary dynamically in both time and space and contribute in the regulation of cellular behavior and fate processes [2]. When developing new in vitro models or functional bio-constructs, the ability of spatially and functionally replicating the native tissue microarchitecture is a crucial step to obtain reliable and physiologically consistent cell responses [3]. Indeed, while traditional 2D in vitro culture techniques lack in reproducing the complexity found in vivo, 3D models have been demonstrated to recapitulate unprecedented cues from the native environment [4].…”

Selective comparison of gelling agents as neural cell culture matrices for long-term microelectrode array electrophysiology

“…Some popular strategies for establishing a more organotypic-like 3D cell culture are the use of rigid scaffolds (Carletti et al, 2011) (e.g., microbeads (Frega et al, 2014) or protein-binding polymer scaffolds (Greiner et al, 2012)) coated with ECM-like adhesion and signaling factors, the direct embedding of cells into detergent-extracted ECM scaffolds (Ott et al, 2008) or functionalized hydrogels. Hydrogels are network-like polymers with high water content (Buwalda et al, 2014).…”