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Updated: Jul 1, 2025
Isolating, Sequencing and Analyzing Extracellular MicroRNAs from Human Mesenchymal Stem Cells
Published on: March 8, 2019
MicroRNA-18a prevents senescence of mesenchymal stem cells by targeting CTDSPL
Bo Sun1, Xian-Hui Meng1, Yu-Min Li1
1State Key Laboratory of Digital Medical Engineering, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.
Abstract:
Stem cell therapy requires massive-scale homogeneous stem cells under strict qualification control. However, Prolonged ex vivo expansion impairs the biological functions and results in senescence of mesenchymal stem cells (MSCs). We investigated the function of CTDSPL in the premature senescence process of MSCs and clarified that miR-18a-5p played a prominent role in preventing senescence of long-term cultured MSCs and promoting the self-renewal ability of MSCs. Over-expression of CTDSPL resulted in an enlarged morphology, up-regulation of p16 and accumulation of SA-β-gal of MSCs. The reduced phosphorylated RB suggested cell cycle arrest of MSCs. All these results implied that CTDSPL induced premature senescence of MSCs. We further demonstrated that miR-18a-5p was a putative regulator of CTDSPL by luciferase reporter assay. Inhibition of miR-18a-5p promoted the expression of CTDSPL and induced premature senescence of MSCs. Continuous overexpression of miR-18a-5p improved self-renewal of MSCs by reducing ROS level, increased expression of Oct4 and Nanog, and promoted growth rate and differentiation capability. We reported for the first time that the dynamic interaction of miR-18a-5p and CTDSPL is crucial for stem cell senescence.
Insights
CTDSPL induces premature senescence in mesenchymal stem cells (MSCs). MicroRNA-18a-5p (miR-18a-5p) prevents this senescence, maintaining MSC self-renewal and function during long-term culture.
Area of Science:
- Stem Cell Biology
- Molecular Biology
- Cellular Senescence
Background:
- Mesenchymal stem cells (MSCs) are crucial for regenerative medicine but undergo senescence during prolonged ex vivo expansion, limiting their therapeutic potential.
- Understanding the molecular mechanisms driving MSC senescence is essential for improving cell therapy protocols.
Purpose of the Study:
- To investigate the role of CTDSPL in the premature senescence of MSCs.
- To elucidate the function of miR-18a-5p in regulating MSC senescence and self-renewal.
Main Methods:
- Overexpression of CTDSPL and miR-18a-5p in MSCs.
- Assessment of senescence markers (morphology, p16, SA-β-gal, cell cycle).
- Luciferase reporter assay to confirm miR-18a-5p regulation of CTDSPL.
- Evaluation of self-renewal markers (ROS levels, Oct4, Nanog, growth rate, differentiation).
Main Results:
- CTDSPL overexpression induced premature senescence in MSCs, characterized by morphological changes, cell cycle arrest, and increased senescence markers.
- miR-18a-5p was identified as a direct regulator of CTDSPL, with its inhibition promoting CTDSPL expression and MSC senescence.
- Overexpression of miR-18a-5p attenuated CTDSPL-induced senescence, reduced ROS levels, enhanced self-renewal markers (Oct4, Nanog), and improved growth and differentiation capabilities.
Conclusions:
- The dynamic interaction between miR-18a-5p and CTDSPL is critical for controlling MSC senescence.
- miR-18a-5p acts as a key protective factor against premature senescence in long-term cultured MSCs, preserving their self-renewal and therapeutic potential.
Related Concept Videos
01:19Mesenchymal Stem Cells
01:22MicroRNAs
02:15Replicative Cell Senescence