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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Single-cell RNA sequencing reveals the heterogeneity of adipose tissue-derived mesenchymal stem cells under chondrogenic induction

  • Jeewan Chun (Department of Oral Microbiology, College of Dentistry, Kyung Hee University) ;
  • Ji-Hoi Moon (Department of Oral Microbiology, College of Dentistry, Kyung Hee University) ;
  • Kyu Hwan Kwack (Department of Oral Microbiology, College of Dentistry, Kyung Hee University) ;
  • Eun-Young Jang (Department of Oral Microbiology, College of Dentistry, Kyung Hee University) ;
  • Saebyeol Lee (Department of Life Science, Chung-Ang University) ;
  • Hak Kyun Kim (Department of Life Science, Chung-Ang University) ;
  • Jae-Hyung Lee (Department of Oral Microbiology, College of Dentistry, Kyung Hee University)
  • Received : 2023.09.01
  • Accepted : 2023.10.12
  • Published : 2024.05.31
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Abstract

This study investigated how adipose tissue-derived mesenchymal stem cells (AT-MSCs) respond to chondrogenic induction using droplet-based single-cell RNA sequencing (scRNA-seq). We analyzed 37,219 high-quality transcripts from control cells and cells induced for 1 week (1W) and 2 weeks (2W). Four distinct cell clusters (0-3), undetectable by bulk analysis, exhibited varying proportions. Cluster 1 dominated in control and 1W cells, whereas clusters (3, 2, and 0) exclusively dominated in control, 1W, and 2W cells, respectively. Furthermore, heterogeneous chondrogenic markers expression within clusters emerged. Gene ontology (GO) enrichment analysis of differentially expressed genes unveiled cluster-specific variations in key biological processes (BP): (1) Cluster 1 exhibited up-regulation of GO-BP terms related to ribosome biogenesis and translational control, crucial for maintaining stem cell properties and homeostasis; (2) Additionally, cluster 1 showed up-regulation of GO-BP terms associated with mitochondrial oxidative metabolism; (3) Cluster 3 displayed up-regulation of GO-BP terms related to cell proliferation; (4) Clusters 0 and 2 demonstrated similar up-regulation of GO-BP terms linked to collagen fibril organization and supramolecular fiber organization. However, only cluster 0 showed a significant decrease in GO-BP terms related to ribosome production, implying a potential correlation between ribosome regulation and the differentiation stages of AT-MSCs. Overall, our findings highlight heterogeneous cell clusters with varying 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.

Keywords

Acknowledgement

This research was supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Science & ICT (NRF-2022R1F1A1071248) and the Chung-Ang University Research Scholarship Grants in 2021.

References

  1. Biehl JK and Russell B (2009) Introduction to stem cell therapy. J Cardiovasc Nurs 24, 98-103
  2. Macrin D, Joseph JP, Pillai AA and Devi A (2017) Eminent sources of adult mesenchymal stem cells and their therapeutic imminence. Stem Cell Rev Rep 13, 741-756
  3. Si Z, Wang X, Sun C et al (2019) Adipose-derived stem cells: sources, potency, and implications for regenerative therapies. Biomed Pharmacother 114, 108765
  4. Zuk PA, Zhu M, Mizuno H et al (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7, 211-228
  5. Fraser JK, Wulur I, Alfonso Z and Hedrick MH (2006) Fat tissue: an underappreciated source of stem cells for biotechnology. Trends Biotechnol 24, 150-154
  6. Kasir R, Vernekar VN and Laurencin CT (2015) Regenerative engineering of cartilage using adipose-derived stem cells. Regen Eng Transl Med 1, 42-49
  7. Pak J, Lee JH, Pak N et al (2018) Cartilage regeneration in humans with adipose tissue-derived stem cells and adipose stromal vascular fraction cells: updated status. Int J Mol Sci 19, 2146
  8. Meng HY, Lu V and Khan W (2021) Adipose tissue-derived mesenchymal stem cells as a potential restorative treatment for cartilage defects: a PRISMA review and meta-analysis. Pharmaceuticals (Basel) 14, 1280
  9. Varghese J, Griffin M, Mosahebi A and Butler P (2017) Systematic review of patient factors affecting adipose stem cell viability and function: implications for regenerative therapy. Stem Cell Res Ther 8, 45
  10. Bunnell BA (2021) Adipose tissue-derived mesenchymal stem cells. Cells 10, 3433
  11. Liu X, Xiang Q, Xu F et al (2019) Single-cell RNA-seq of cultured human adipose-derived mesenchymal stem cells. Sci Data 6, 190031
  12. Yang S, Cho Y and Jang J (2021) Single cell heterogeneity in human pluripotent stem cells. BMB Rep 54, 505-515
  13. Ziegenhain C, Vieth B, Parekh S et al (2017) Comparative analysis of single-cell RNA sequencing methods. Mol Cell 65, 631-643
  14. Zhou WM, Yan YY, Guo QR et al (2021) Microfluidics applications for high-throughput single cell sequencing. J Nanobiotechnology 19, 312
  15. Macosko EZ, Basu A, Satija R et al (2015) Highly parallel genome-wide expression profiling of individual cells using nanoliter droplets. Cell 161, 1202-1214
  16. Stromps JP, Paul NE, Rath B, Nourbakhsh M, Bernhagen J and Pallua N (2014) Chondrogenic differentiation of human adipose-derived stem cells: a new path in articular cartilage defect management? Biomed Res Int 2014, 740926
  17. Xu J, Wang W, Ludeman M et al (2008) Chondrogenic differentiation of human mesenchymal stem cells in three-dimensional alginate gels. Tissue Eng Part A 14, 667-680
  18. Ilicic T, Kim JK, Kolodziejczyk AA et al (2016) Classification of low quality cells from single-cell RNA-seq data. Genome Biol 17, 29
  19. Bock FJ and Tait SWG (2020) Mitochondria as multifaceted regulators of cell death. Nat Rev Mol Cell Biol 21, 85-100
  20. Singh AM (2015) Cell cycle-driven heterogeneity: on the road to demystifying the transitions between "poised" and "restricted" pluripotent cell states. Stem Cells Int 2015, 219514
  21. Tirosh I, Izar B, Prakadan SM et al (2016) Dissecting the multicellular ecosystem of metastatic melanoma by single-cell RNA-seq. Science 352, 189-196
  22. Hou W, Duan L, Huang C et al (2021) Cross-tissue characterization of heterogeneities of mesenchymal stem cells and their differentiation potentials. Front Cell Dev Biol 9, 781021
  23. Leung VY, Gao B, Leung KK et al (2011) SOX9 governs differentiation stage-specific gene expression in growth plate chondrocytes via direct concomitant transactivation and repression. PLoS Genet 7, e1002356
  24. Ruijtenberg S and van den Heuvel S (2016) Coordinating cell proliferation and differentiation: antagonism between cell cycle regulators and cell type-specific gene expression. Cell Cycle 15, 196-212
  25. Gabut M, Bourdelais F and Durand S (2020) Ribosome and translational control in stem cells. Cells 9, 497
  26. Yan W, Diao S and Fan Z (2021) The role and mechanism of mitochondrial functions and energy metabolism in the function regulation of the mesenchymal stem cells. Stem Cell Res Ther 12, 140
  27. Nolfi-Donegan D, Braganza A and Shiva S (2020) Mitochondrial electron transport chain: oxidative phosphorylation, oxidant production, and methods of measurement. Redox Biol 37, 101674
  28. Zhang Y, Marsboom G, Toth PT and Rehman J (2013) Mitochondrial respiration regulates adipogenic differentiation of human mesenchymal stem cells. PLoS One 8, e77077
  29. Shum LC, White NS, Mills BN, Bentley KL and Eliseev RA (2016) Energy metabolism in mesenchymal stem cells during osteogenic differentiation. Stem Cells Dev 25, 114-122