Genomic analysis of dental pulp mesenchymal stromal cells for therapeutic applications
Kunio Hirai1,2, Yuko Nitahara-Kasahara1, Yuko Okamoto1
1Division of Molecular and Medical Genetics, Center for Gene and Cell Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
Introduction:
Changes in cell characteristics and the accumulation of gene mutations during expansion culture present significant challenges for the clinical translation of stem cell transplantation technologies. Reduced cell quality may lead to diminished therapeutic efficacy and unexpected cellular dysfunction after transplantation. Mesenchymal stromal cells (MSCs), including dental pulp stromal cells (DPSCs), are promising candidates for clinical application because of their multipotent differentiation capacity, immunosuppressive properties, and proliferative potential. However, robust strategies to assess genomic stability during large-scale cell preparation remain insufficient.
Methods:
DPSCs derived from healthy donors maintained safety profiles and exhibited delayed senescence even after more than 10 passages under large-scale expansion conditions. To evaluate genomic stability, DPSCs derived from healthy individuals and patients with type 1 diabetes mellitus were analyzed after successive passages. Copy number variations (CNVs) were assessed using CytoScan, and disease-related gene mutations were analyzed using the TruSight One Sequencing Panel.
Results:
During long-term expansion, increases in chromosomal CNVs and de novo gene mutations were observed at earlier stages in cells showing reduced proliferative capacity, depending on donor characteristics. We identified mutation accumulation patterns and genes that were particularly susceptible to substitutions or insertions/deletions during expansion, indicating inter-donor variability in genomic alterations in DPSCs.
Conclusion:
This work addresses a critical bottleneck in stem cell therapy by integrating genomic risk profiling into early-stage cell quality assessment, enabling safer and more reliable cell product development. Furthermore, our findings highlight the feasibility of applying medical genomic tools to preemptively identify putative high-risk cell populations during stem cell manufacturing, thereby providing a practical framework for improving the safety and consistency of cell-based transplantation therapies.
