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Integrated multi-assay culture model for stem cell chondrogenic differentiation

Prosser, Amy; Scotchford, Colin; Roberts, George; Grant, David; Sottile, Virginie

Authors

Amy Prosser

George Roberts

DAVID GRANT david.grant@nottingham.ac.uk
Professor of Materials Science

Virginie Sottile



Abstract

Recent osteochondral repair strategies highlight the promise of mesenchymal progenitors, an accessible stem cell source with osteogenic and chondrogenic potential, used in conjunction with biomaterials for tissue engineering. For this, regenerative medicine approaches require robust models to ensure selected cell populations can generate the desired cell type in a reproducible and measurable manner. Techniques for in vitro chondrogenic differentiation are well-established but largely qualitative, relying on sample staining and imaging. To facilitate the in vitro screening of pro-chondrogenic treatments, a 3D micropellet culture combined with three quantitative GAG assays has been developed, with a fourth parallel assay measuring sample content to enable normalisation. The effect of Transforming Growth Factor beta (TGF-β) used to validate this culture format produced a measurable increase in proteoglycan production in the parallel assays, in both 2D and 3D culture configurations. When compared to traditional micropellets, the monolayer format appeared less able to detect changes in cell differentiation, however in-well 3D cultures displayed a significant differential response. Effects on collagen 2 expression confirmed these observations. Based on these results, a microplate format was optimised for 3D culture, in a high-throughput in-well configuration. This model showed improved sensitivity and confirmed the 3D micropellet in-well quantitative assays as an effective differentiation format compatible with streamlined, high-throughput chondrogenic screens.

Journal Article Type Article
Publication Date Feb 22, 2019
Journal International Journal of Molecular Sciences
Print ISSN 1661-6596
Electronic ISSN 1422-0067
Publisher MDPI
Peer Reviewed Peer Reviewed
Volume 20
Issue 4
Article Number 951
APA6 Citation Prosser, A., Scotchford, C., Roberts, G., Grant, D., & Sottile, V. (2019). Integrated multi-assay culture model for stem cell chondrogenic differentiation. International Journal of Molecular Sciences, 20(4), https://doi.org/10.3390/ijms20040951
DOI https://doi.org/10.3390/ijms20040951
Keywords Stem cell differentiation; 3D culture; Chondrogenesis; Multimodal analysis; Quantitative assay
Publisher URL https://www.mdpi.com/1422-0067/20/4/951

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