Abstract:
:Scaffold-free 3D cell cultures (e.g. pellet cultures) are widely used in medical science, including cartilage regeneration. Their drawbacks are high time/reagent consumption and lack of early readout parameters. While optimisation was achieved by automation or simplified spheroid generation, most culture systems remain expensive or require tedious procedures. The aim of this study was to establish a system for resource efficient spheroid generation. This was achieved by compartmentation of cell culture surfaces utilising laser engraving (grid plates). This compartmentation triggered autonomous spheroid formation via rolling-up of the cell monolayer in human adipose-derived stem cells (ASC/TERT1) and human articular chondrocytes (hAC)-ASC/TERT1 co-cultures, when cultivated on grid plates under chondrogenic conditions. Plates with 3 mm grid size yielded stable diameters (about 300 μm). ASC/TERT1 spheroids fully formed within 3 weeks while co-cultures took 1-2 weeks, forming significantly faster with increasing hAC ratio (p<0.05 and 0.01 for 1:1 and 1:4 ASC/TERT1:hAC ratio respectively). Co-cultures showed slightly lower spheroid diameter, due to earlier spheroid formation and incomplete monolayer formation. However, this was associated with more regular matrix distribution in the co-culture. Both showed differentiation capacity comparable to standard pellet culture in (immune-)histochemistry and RT-qPCR. To assess usability for cartilage repair, spheroids were embedded into a hydrogel (fibrin), yielding cellular outgrowth and matrix deposition, which was especially pronounced in co-cultures. The herein presented novel cell culture system is not only a promising tool for autonomous spheroid generation with the potential of experimental and clinical application in tissue engineering but also for high-throughput analysis for both pharmaceutical and therapeutic uses.
journal_name
Biofabricationjournal_title
Biofabricationauthors
Fürsatz M,Gerges P,Wolbank S,Nürnberger Sdoi
10.1088/1758-5090/abe186subject
Has Abstractpub_date
2021-01-29 00:00:00eissn
1758-5082issn
1758-5090pub_type
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