AccScience Publishing / IJB / Online First / DOI: 10.36922/IJB025290291
RESEARCH ARTICLE
Early Access

Automated and scalable fabrication of scaffolded spheroids as building blocks for modular tissue engineering

Gregor Weisgrab1,2 Sebastian Rudi Adam Kratz3 Mario Rothbauer3,4 Martin Frauenlob3 Peter Ertl3 Aleksandr Ovsianikov1,2*
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1 Research Group 3D Printing and Biofabrication, Institute of Materials Science and Technology, TU Wien, Getreidemarkt 9/308, 1060 Vienna, Austria
2 Austrian Cluster for Tissue Regeneration, Austrian Cluster for Tissue Regeneration
3 Cell Chip Group, Institute of Applied Synthetic Chemistry, Institute of Chemical Technologies and Analytics, TU Wien, Getreidemarkt 9/163, 1060 Vienna, Austria
4 Karl Chiari Lab for Orthopedic Biology, Department of Orthopedics and Trauma Surgery, Medical University of Vienna, Waehringerguertel 18-20, 1090 Vienna, Austria
Received: 16 July 2025 | Accepted: 11 September 2025 | Published online: 11 September 2025
© 2025 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Abstract

Current tissue engineering approaches for most part utilize either a scaffold-based or a scaffold-free strategies. Recently, a third strategy demonstrated a synergetic method leveraging the benefits of both approaches without their typical disadvantages. For this, spheroids are formed within highly porous microscaffolds that then act as building blocks for larger tissue constructs produced by self-assembly. The work presented here demonstrates the fabrication of a large number of such building blocks in an automated way, tested in up to 1536-well plates. We first developed a microfluidic device capable of sorting highly porous microscaffolds with a diameter of 300 µm at high flow speeds of up to 300 mm/s. A fluorescent setup was developed to single out only intact microscaffolds and deposit them one-by-one in separate wells of cell culture plates. Subsequently, the system automatically dispensed culture medium with suspended human adipose-derived stem cells into each well. Spheroids were formed within 24 hours of culture with a formation efficiency of 95%. This study reports the development of an automated microfluidic device capable of depositing single microscaffolds and the cells required to form scaffolded spheroids, thereby enabling the benefits of the third strategy of tissue engineering at scale fit for screening applications and clinical translation.

Keywords
High-resolution 3D printing
Tissue engineering
Microfluidic automation
Scale-up production
Microscaffolds
Funding
This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (Grant agreement No. 772464).
Conflict of interest
The authors declare they have no competing interests
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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing