Human mesenchymal stem cell expression program upon extended ex-vivo cultivation, as revealed by 2-DE-based...

Area of Science:

• Cell Biology
• Proteomics
• Biotechnology

Background:

• Human mesenchymal stem cells (MSC) are crucial for cellular therapy due to their differentiation and immunomodulatory potential.
• Ex-vivo expansion is necessary for clinical applications but leads to culture-induced senescence, reducing cell function.
• Understanding senescence mechanisms is vital for ensuring the efficacy and safety of expanded MSC.

Purpose of the Study:

• To investigate the molecular mechanisms of cellular senescence in bone marrow MSC during extended ex-vivo expansion.
• To compare the proteome profiles of early-passage (P3) and late-passage (P7) MSC to identify changes associated with senescence.

Main Methods:

• Quantitative proteomics using two-dimensional gel electrophoresis (2-DE) was employed.
• Protein expression profiles of Passage 3 (P3) and Passage 7 (P7) human bone marrow MSC were compared.

Main Results:

• Senescent P7 MSC showed decreased abundance of structural/cytoskeleton and folding/stress response proteins.
• Increased abundance of proteins related to energy metabolism, cell cycle regulation, aging, and apoptosis was observed in P7 MSC.
• Significant alterations in post-transcriptional modifications, particularly for cytoskeleton components like β-actin and vimentin, were identified.

Conclusions:

• Ex-vivo expansion-induced senescence in MSC is linked to impaired cytoskeleton remodeling and protein repair mechanisms.
• Proteomic analysis provides mechanistic insights into reduced proliferative and clonogenic potential of aged MSC.
• Proteome profiling can serve as a quality control strategy for clinical-grade expanded MSC.