Bioassemblying Macro-Scale, Lumnized Airway Tubes of Defined Shape via Multi-Organoid Patterning and Fusion

Abstract

Epithelial, stem-cell derived organoids are ideal building blocks for tissue engineering, however, scalable and shape-controlled bioassembly of epithelial organoids into larger and anatomical structures has yet to be achieved. Here, a robust organoid engineering approach, Multi-Organoid Patterning and Fusion (MOrPF), is presented to assemble individual airway organoids of different sizes into upscaled, scaffold-free airway tubes with pre-defined shapes. Multi-Organoid Aggregates (MOAs) undergo accelerated fusion in a matrix-depleted, free-floating environment, possess a continuous lumen and maintain prescribed shapes without an exogenous scaffold interface. MOAs in the floating culture exhibit a well-defined three-stage process of inter-organoid surface integration, luminal material clearance and lumina connection. The observed shape stability of patterned MOAs is confirmed by theoretical modelling based on organoid morphology and the physical forces involved in organoid fusion. Immunofluorescent characterization shows that fused MOA tubes possess an unstratified epithelium consisting mainly of tracheal basal stem cells. By generating large, shape-controllable organ tubes, MOrPF enables upscaled organoid engineering towards integrated organoid-devices and structurally complex organ tubes.

Volumetric Bioprinting of Organoids and Optically Tuned Hydrogels to Build Liver‐Like Metabolic Biofactories

“…To date, organoid shaping via bioprinting has been demonstrated via extrusion of a suspension of single stem cells, which are then led to re‐form into organoids post‐printing. [ 50 ] Alternatively, biofabrication of pre‐generated organoids has been prevalently performed via molding, [ 51 ] individual spheroid dispensing, [ 5 ] or robotic‐assisted pick‐and‐place techniques. [ 52 ] Although yielding impressive results in terms of generating tissues with high cell content, these approaches are limited to relatively simple 3D geometries, and rely on the printing of thick filaments/spheroids with a 400–1000 µm diameter range to achieve simple tubular structures.…”

“…[ 52 ] Although yielding impressive results in terms of generating tissues with high cell content, these approaches are limited to relatively simple 3D geometries, and rely on the printing of thick filaments/spheroids with a 400–1000 µm diameter range to achieve simple tubular structures. [ 5,50–52 ] Complementing the possibilities granted by such strategies, the ability of VBP to print pristine, undamaged organoids offers an alternative to facilitate the free‐form generation of intact organoid‐laden constructs. Printing morphologically intact organoids can be advantageous for applications aiming to preserve the organoid pre‐deposited ECM, given the increasing evidence that cells embedded in biomaterials alter their behavior via contact with the nascent, self‐synthesized ECM.…”

Mammary Tumor Organoid Culture in Non‐Adhesive Alginate for Luminal Mechanics and High‐Throughput Drug Screening

“…Organoid lumen formation arises from the interplay of mechanical and biochemical signaling. [ 30 ] Measuring biomechanics, [ 31 ] especially the luminal mechanics, [ 26 ] has been the subject of intense scholarly debate. Various methods have been established to measure the luminal pressure, such as the micropressure probe, gel deformation assay, pressure sensors, deformable beads, traction microscopy, and atomic force microscopy.…”

Advancing Tissue Decellularized Hydrogels for Engineering Human Organoids

“…Reproduced under the terms of the Creative Commons CC BY license. [ 172 ] Copyright 2021, Ye Liu et al D) Representative images of GFP‐positive MSCs delivered in: D1) saline solution and D2) ECM pre‐gel solution. Arrows point to GFP‐positive cells.…”

“…A comprehensive investigation of MOAs’ culture conditions and niche engineering through the use of a precise dECM composition and timing of addition of key biomolecular elements could improve the maintenance of cells’ polarity and differentiation. [ 172 ] Whereas most lung organoids are established in BME (i.e., Matrigel), the proteins that comprise this matrix are not equal to those present in native lung tissues. While BME comprises mostly collagen type IV and laminin, [ 173 ] the lung matrix has a high content of collagen type I and III and elastin.…”