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Biotechnol Bioeng. 2015 Jul;112(7):1457-71. doi: 10.1002/bit.25557. Epub 2015 Mar 10.

Fabrication of multi-well chips for spheroid cultures and implantable constructs through rapid prototyping techniques.

Author information

1
Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Orthopaedic Institute, Via R. Galeazzi 4, 20161, Milan, Italy.
2
Department of Electronics, Information, and Bioengineering, Politecnico di Milano, Milan, 20133, Italy.
3
Tissue Regeneration Department, University of Twente, 7522 NB, Enschede, The Netherlands.
4
IRCCS Galeazzi Orthopaedic Institute, Milan, 20161, Italy.
5
Department of Complex Tissue Regeneration, Maastricht University, 6200 MD, Maastricht, The Netherlands.
6
Department of Biomedical Sciences for Health, Università degli Studi di Milano, Milan, 20161, Italy.
7
Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Orthopaedic Institute, Via R. Galeazzi 4, 20161, Milan, Italy. matteo.moretti@grupposandonato.it.

Abstract

Three-dimensional (3D) culture models are widely used in basic and translational research. In this study, to generate and culture multiple 3D cell spheroids, we exploited laser ablation and replica molding for the fabrication of polydimethylsiloxane (PDMS) multi-well chips, which were validated using articular chondrocytes (ACs). Multi-well ACs spheroids were comparable or superior to standard spheroids, as revealed by glycosaminoglycan and type-II collagen deposition. Moreover, the use of our multi-well chips significantly reduced the operation time for cell seeding and medium refresh. Exploiting a similar approach, we used clinical-grade fibrin to generate implantable multi-well constructs allowing for the precise distribution of multiple cell types. Multi-well fibrin constructs were seeded with ACs generating high cell density regions, as shown by histology and cell fluorescent staining. Multi-well constructs were compared to standard constructs with homogeneously distributed ACs. After 7 days in vitro, expression of SOX9, ACAN, COL2A1, and COMP was increased in both constructs, with multi-well constructs expressing significantly higher levels of chondrogenic genes than standard constructs. After 5 weeks in vivo, we found that despite a dramatic size reduction, the cell distribution pattern was maintained and glycosaminoglycan content per wet weight was significantly increased respect to pre-implantation samples. In conclusion, multi-well chips for the generation and culture of multiple cell spheroids can be fabricated by low-cost rapid prototyping techniques. Furthermore, these techniques can be used to generate implantable constructs with defined architecture and controlled cell distribution, allowing for in vitro and in vivo investigation of cell interactions in a 3D environment.

KEYWORDS:

3D model; cell spheroid; implantable scaffold; rapid prototyping; tissue engineering

PMID:
25678107
DOI:
10.1002/bit.25557
[Indexed for MEDLINE]

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