Integrated Multi-Assay Culture Model for Stem Cell Chondrogenic Differentiation

International Journal of Molecular Sciences
Amy ProsserV 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 obs...Continue Reading

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Citations

Oct 5, 2019·Scientific Reports·Manuela MinguzziRosa Maria Borzì
Apr 24, 2020·Journal of Biomedical Materials Research. Part a·Yu-Hsun ChangDah-Ching Ding
Jan 21, 2021·Journal of Functional Morphology and Kinesiology·Graziana MonacoMartin J Stoddart

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Methods Mentioned

BETA
dot blotting
ELISA
Assay

Software Mentioned

ImageJ
GraphPad Prism

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