Next-generation approaches for mesenchymal stem cell characterization and therapeutic application

Mesenchymal stem cells (MSCs) have been the center of focus in the field of regenerative medicine owing to their intrinsic multipotency, ability for immunomodulation, and paracrine-mediated recovery processes. In spite of extensive research in the clinical setting, the therapeutic application of MSCs is still hindered by challenges from intrinsic heterogeneity, donor-to-donor variability, senescence of cells, and variability in potency. In this review, the recent biological understanding of MSCs is stringently evaluated concerning their role in immunomodulation, anti-apoptotic function, anti-fibrotic activity, angiogenesis, mitochondrial transfer, and antioxidant function. Additionally, it highlights the therapeutic potential of MSCs in osteoarthritis, cardiovascular disorders, and several neuropsychiatric, immunological, and dermatological ailments. Emphasis is placed on innovative approaches being generated for eroding the drawbacks of MSCs, which involve utilizing single-cell and spatial transcriptomics for deconvoluting cellular heterogeneity, applying CRISPR/Cas9 gene editing for genetic modification, and utilizing bioengineering approaches like 3D bioprinting and organ-on-a-chip systems. In addition, cell-free therapy from MSC-derived exosomes and secretomes is discussed as a novel substitute for conventional cell-based therapy, as they pose improved security, stability, and scalability. The integration of preconditioning, gene editing, and targeted delivery is also discussed as a strategic mechanism for enhanced therapeutic efficacy. In all, the review provides a well-integrated framework for MSC-based treatment mechanisms, clinical indications, and technological innovations, furthering the advancement of regenerative medicine.