Trehalose enhances neuronal differentiation with VEGF secretion in human iPSC-derived neural stem/progenitor cells.

INTRODUCTION

Cell transplantation therapy has emerged as a promising approach in regenerative medicine for treating neurological diseases. Neural stem/progenitor cell (NS/PC) transplantation has demonstrated therapeutic efficacy; however, its potential remains limited by suboptimal differentiation and insufficient secretion of pro-healing growth factors. Trehalose, a glucose disaccharide, has been shown to exert neuroprotective effects by inducing autophagy and stabilizing cellular structures. Recent studies suggest that trehalose can modulate growth factor secretion through the CDKN1A/p21 pathway. However, its impact on human induced pluripotent stem cell-derived NS/PCs (hiPSC-NS/PCs) remains unclear. This study investigates the effect of trehalose on neuronal differentiation, cell viability, and growth factor expression in hiPSC-NS/PCs to explore its potential in enhancing transplantation therapy.

METHODS

hiPSC-NS/PCs were cultured as neurospheres and treated with trehalose (10 mg/ml or 40 mg/ml) for 7 days. Cell viability was assessed using CellTiter Glo® assay. Gene expression analysis was conducted via qRT-PCR and RNA-seq, particularly focusing on , , , and . Protein expression of SOX2 was analyzed via western blotting. Neurite outgrowth was evaluated using MAP2 immunostaining following differentiation. Statistical significance was set at p < 0.05.

RESULTS

Treatment with 10 mg/ml trehalose upregulated expression and promoted neuronal differentiation, as evidenced by reduced SOX2 expression and enhanced neurite outgrowth. RNA-seq analysis revealed the activation of growth factor-related pathways, including upregulation, which persisted even after trehalose withdrawal (p = 0.016). However, high concentration (40 mg/ml) significantly reduced cell viability (p = 0.032), suggesting dose-dependent cytotoxicity.

CONCLUSION

Trehalose enhances neuronal differentiation and secretion in hiPSC-NS/PCs, potentially augmenting the efficacy of transplantation therapy. These findings suggest that trehalose may serve as a valuable adjunct for neural regeneration, though optimal dosing must be determined to balance differentiation enhancement and cell viability. Further studies are warranted to validate its clinical applicability.