Precision 3D‐Printed Cell Scaffolds Mimicking Native Tissue Composition and Mechanics

Abstract

Cellular dynamics are modeled by the 3D architecture and mechanics of the extracellular matrix (ECM) and vice versa. These bidirectional cell‐ECM interactions are the basis for all vital tissues, many of which have been investigated in 2D environments over the last decades. Experimental approaches to mimic in vivo cell niches in 3D with the highest biological conformity and resolution can enable new insights into these cell‐ECM interactions including proliferation, differentiation, migration, and invasion assays. Here, two‐photon stereolithography is adopted to print up to mm‐sized high‐precision 3D cell scaffolds at micrometer resolution with defined mechanical properties from protein‐based resins, such as bovine serum albumin or gelatin methacryloyl. By modifying the manufacturing process including two‐pass printing or post‐print crosslinking, high precision scaffolds with varying Young's moduli ranging from 7‐300 kPa are printed and quantified through atomic force microscopy. The impact of varying scaffold topographies on the dynamics of colonizing cells is observed using mouse myoblast cells and a 3D‐lung microtissue replica colonized with primary human lung fibroblast. This approach will allow for a systematic investigation of single‐cell and tissue dynamics in response to defined mechanical and bio‐molecular cues and is ultimately scalable to full organs.

Single Cell Bioprinting with Ultrashort Laser Pulses

“…This is highlighted in Figure 3b, where individual hMSCs were placed one by one in a premanufactured cross-linked bovine serum albumin (BSA) scaffold. [57,58] Note that after the third cell was transferred to the BSA scaffold and placed in close proximity to the first and the second transferred cell, the second cell was slightly shifted. This was most likely driven by a hydrodynamic flow on the scaffold's surface, caused by the hydrogel carrying the third cell.…”

“…BSA resin preparation and high‐resolution 3D printing were conducted as described elsewhere. [ 57 ] In brief, 40 wt% BSA stock solution was mixed with 85 m m rose bengal stock solution in the ratio 9:1. A droplet of this suspension was placed on a 170 µm thick glass slide.…”

Local Membrane Curvature Pins and Guides Excitable Membrane Waves in Chemotactic and Macropinocytic Cells - Biomedical Insights From an Innovative Simple Model

“…This crowding may affect compartmentalization and other mechanisms in living cells that allow proteins to sense, stabilize or generate a high local membrane curvature (McMahon and Boucrot, 2015 ), such as PTEN, a cytosolic protein that associates with the plasma membrane. Further experimental investigations on precisely controlled curvatures of artificial cell membrane systems using microfabrication (Komiya et al, 2020 ) and high-precision 3D printing (Erben et al, 2020 ) may enhance our understanding of these findings.…”

Breathing in vitro: Designs and applications of engineered lung models

“…However, work still needs to be performed in order to have a coherent approximation of scalable lung-specific design. With the goal of obtaining in vivo-like structures, also Erben et al 228 used stereolithography to print mm-sized high precision 3D scaffolds at micrometer resolution.…”