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Development of gene expression signatures for bone marrow mesenchymal stem cells with 3D culture in collagen hydrogel

ABSTRACT: To investigate the mechanisms underlying chondrocyte differentiation of bone marrow mesenchymal stem cells during 3D culture, we employed whole genome microarray expression profiling as a discovery platform to identify key genes influencing differentiation. By comparing differentially expressed genes at different time points, we identified POU5F1 and several autophagy-related genes as potential key players. By using PCR technology and autophagy-related assays, we confirmed that POU5F1 and autophagy pathways influence the chondrogenic differentiation of bone marrow-derived mesenchymal stem cells.

ORGANISM(S): Oryctolagus cuniculus

PROVIDER: GSE315306 | GEO | 2026/01/07

REPOSITORIES: GEO

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Project description:Fibroblast growth factor-2 delays the loss of chondrogenic potential in adult bone marrow-derived mesenchymal stem cells; We compared human mesenchymal stem cells (hMSCs), expanded long-term with and without fibroblast growth factor (FGF) supplementation, with respect to their proliferation rate, and ability to differentiate along the chondrogenic pathway in vitro. hMSCs expanded in FGF-supplemented medium proliferated more rapidly than those expanded under control conditions. Aggregates of FGF-treated cells exhibited chondrogenic differentiation at passages 1 through 7, although, in some of the preparations, chondrogenic differentiation was somewhat diminished after seventh passage. Aggregates made with control cells differentiated along the chondrogenic lineage at first passage but exhibited only marginal chondrogenic differentiation after four passages and failed to form cartilage after seven passages. Microarray analysis of gene expression identified 334 transcripts that were differentially expressed in fourth passage control cells which had reduced chondrogenic potential compared to fourth passage FGF-treated cells which retained this differentiation capacity and 243 transcripts that were differentially expressed when comparing them to first passage control cells which were also capable of differentiating into chondrocytes. The intersection of these analyses yielded 49 transcripts that were differentially expressed in cells that exhibited chondrogenic differentiation in vitro compared to cells that did not. These preliminary data must now be validated to verify whether the different gene expression profiles translate into functional differences. These findings suggest that care should be exercised when extensively expanding these cells for cartilage tissue engineering applications. Experiment Overall Design: hMSCs from three different donors were expanded for up to seven passages with or without FGF supplementation. At the end of first, fourth and seventh passages the chondrogenic potential and gene expression progfiles were analyzed.

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Development of gene expression signatures for bone marrow mesenchymal stem cells with 3D culture in collagen hydrogel

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