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Semi-Automated Phenotypic Analysis of Functional 3D Spheroid Cell Cultures
Published on: August 18, 2023
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Autonomous spheroid formation by culture plate compartmentation
Marian Fürsatz1,2, Peter Gerges2,3, Susanne Wolbank2,4
1Department of Orthopedics and Trauma-Surgery, Division of Trauma-Surgery, Medical University of Vienna, Vienna, Austria.
Biofabrication
|January 29, 2021
Summary
This study introduces a novel, resource-efficient method using laser-engraved grid plates for generating scaffold-free 3D cell spheroids, enabling faster formation and early readouts for tissue engineering applications.
Area of Science:
- Biotechnology
- Tissue Engineering
- Cell Biology
Background:
- Scaffold-free 3D cell cultures, like pellet cultures, are vital for medical science, particularly cartilage regeneration.
- Existing methods face challenges including high costs, time consumption, reagent usage, and a lack of early readout parameters.
- Previous optimizations using automation or simplified spheroid generation often remain expensive or complex.
Purpose of the Study:
- To develop a resource-efficient system for generating 3D cell spheroids.
- To incorporate early readout parameters into the spheroid culture system.
- To establish a novel method for autonomous spheroid generation suitable for tissue engineering and biofabrication.
Main Methods:
- Utilized laser engraving to create grid plates for compartmenting cell culture surfaces.
- Induced spheroid formation through self-assembly, contraction, and rolling up of cell monolayers.
- Cultivated human adipose-derived stem cells (ASC/TERT1) and human articular chondrocytes (hACs)-ASC/TERT1 co-cultures under chondrogenic conditions on grid plates.
- Analyzed spheroid formation time, diameter, matrix distribution, differentiation capacity, and post-embedding behavior.
Main Results:
- The grid plate system successfully generated stable spheroids with diameters of approximately 140 µm (1 mm grid) and 300 µm (3 mm grid).
- ASC/TERT1 spheroids formed within 3 weeks; co-cultures formed significantly faster (1-2 weeks), with speed increasing with higher hAC ratios (p<0.05 and p<0.01).
- Co-cultures exhibited more homogeneous matrix distribution and comparable differentiation capacity to standard pellet cultures; they also showed enhanced cellular outgrowth and matrix deposition when embedded in fibrin hydrogel.
Conclusions:
- The novel laser-engraved grid plate system enables autonomous and resource-efficient spheroid generation.
- This method provides early readout parameters and is suitable for applications in tissue engineering and biofabrication, such as creating building blocks for bioprinting.
- The co-culture system demonstrates particular promise for cartilage repair applications due to enhanced matrix deposition and cellular activity.
