Single-cell RNA sequencing reveals the heterogeneity of adipose tissue-derived mesenchymal stem cells under chondrogenic induction
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This study investigated how adipose tissue-derived mesenchymal stem cells (AT-MSCs) respond to chondrogenic inductionusing droplet-based single-cell RNA sequencing (scRNA-seq). We analyzed 37,219 high-quality transcripts from control cellsand cells induced for 1 week (1W) and 2 weeks (2W). Fourdistinct cell clusters (0-3), undetectable by bulk analysis, exhibited varying proportions. Cluster 1 dominated in control and1W cells, whereas clusters (3, 2, and 0) exclusively dominatedin control, 1W, and 2W cells, respectively. Furthermore, heterogeneous chondrogenic markers expression within clusters emerged. Gene ontology (GO) enrichment analysis of differentiallyexpressed genes unveiled cluster-specific variations in key biological processes (BP): (1) Cluster 1 exhibited up-regulation ofGO-BP terms related to ribosome biogenesis and translationalcontrol, crucial for maintaining stem cell properties and homeostasis; (2) Additionally, cluster 1 showed up-regulation of GO-BPterms associated with mitochondrial oxidative metabolism; (3)Cluster 3 displayed up-regulation of GO-BP terms related to cellproliferation; (4) Clusters 0 and 2 demonstrated similar up-regulation of GO-BP terms linked to collagen fibril organizationand supramolecular fiber organization. However, only cluster0 showed a significant decrease in GO-BP terms related toribosome production, implying a potential correlation betweenribosome regulation and the differentiation stages of AT-MSCs. Overall, our findings highlight heterogeneous cell clusters withvarying balances between proliferation and differentiation before,and after, chondrogenic stimulation. This provides enhanced insights into the single-cell dynamics of AT-MSCs during chondrogenic differentiation.
Abstract
This study investigated how adipose tissue-derived mesenchymal stem cells (AT-MSCs) respond to chondrogenic inductionusing droplet-based single-cell RNA sequencing (scRNA-seq). We analyzed 37,219 high-quality transcripts from control cellsand cells induced for 1 week (1W) and 2 weeks (2W). Fourdistinct cell clusters (0-3), undetectable by bulk analysis, exhibited varying proportions. Cluster 1 dominated in control and1W cells, whereas clusters (3, 2, and 0) exclusively dominatedin control, 1W, and 2W cells, respectively. Furthermore, heterogeneous chondrogenic markers expression within clusters emerged. Gene ontology (GO) enrichment analysis of differentiallyexpressed genes unveiled cluster-specific variations in key biological processes (BP): (1) Cluster 1 exhibited up-regulation ofGO-BP terms related to ribosome biogenesis and translationalcontrol, crucial for maintaining stem cell properties and homeostasis; (2) Additionally, cluster 1 showed up-regulation of GO-BPterms associated with mitochondrial oxidative metabolism; (3)Cluster 3 displayed up-regulation of GO-BP terms related to cellproliferation; (4) Clusters 0 and 2 demonstrated similar up-regulation of GO-BP terms linked to collagen fibril organizationand supramolecular fiber organization. However, only cluster0 showed a significant decrease in GO-BP terms related toribosome production, implying a potential correlation betweenribosome regulation and the differentiation stages of AT-MSCs. Overall, our findings highlight heterogeneous cell clusters withvarying balances between proliferation and differentiation before,and after, chondrogenic stimulation. This provides enhanced insights into the single-cell dynamics of AT-MSCs during chondrogenic differentiation.
참고문헌 (29)
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