Gestational age–associated differences in human umbilical cord mesenchymal stem cells: biological, paracrine, and neuroprotective properties

Abstract

Abstract Background Human umbilical cord mesenchymal stem cells (UCMSCs) have shown considerable therapeutic promise for neonatal brain injury. However, whether gestational age influences UCMSC biological characteristics and therapeutic efficacy remains unclear. This study investigated the potential functional and molecular differences between UCMSCs derived from preterm and full-term infants to inform future autologous and allogeneic cell-based therapies. Methods UCMSCs were isolated from full-term and preterm umbilical cords, with preterm samples further classified into uncomplicated preterm (N-P), gestational diabetes exposure (P-GD), and severe preeclampsia exposure (P-SE) groups. MSC identity, viability, proliferation, apoptosis, trilineage differentiation, and extracellular vesicle (EV) characteristics were evaluated. Transcriptomic analysis was performed to compare N-P and full-term (N-F) UCMSCs, followed by RT-PCR validation of key hub genes. Therapeutic efficacy was assessed in a neonatal germinal matrix hemorrhage (GMH) mouse model following intranasal administration of N-F- or N-P-derived UCMSCs, with neurological function and brain injury evaluated by behavioral testing and T2-weighted MRI. Results All UCMSCs met standard MSC characterixation criteria. Compared with N-F UCMSCs, N-P UCMSCs showed increased cell viability as assessed by CCK-8, enhanced chondrogenic differentiation potential, and increased secretion of EVs expressing CD9, CD63, and CD81. In contrast, UCMSCs from the P-GD and P-SE groups displayed reduced proliferative and differentiation capacities. Transcriptomic analysis identified 631 differentially expressed genes between N-P and N-F UCMSCs, with significant enrichment of pathways involved in cell proliferation, differentiation, and EV biology. Network analysis identified CTNNB1 and SRC as key hub genes, and RT-qPCR confirmed differential expression of representative hub genes. In the neonatal GMH model, both N-F- and N-P-derived UCMSCs significantly improved behavioral outcomes and reduced brain injury compared with untreated GMH mice. Although N-P-derived UCMSCs showed a trend toward greater therapeutic benefit, no statistically significant difference was observed between the two treatment groups. Conclusion Healthy preterm UCMSCs display distinct biological and paracrine-related properties while retaining neuroprotective effects comparable to that of full-term UCMSCs. In contrast, adverse intrauterine conditions, including gestational diabetes and severe preeclampsia, are associated with impaired UCMSC biological properties. These findings provide new insights into the developmental regulation of UCMSC function and support further investigation of preterm UCMSCs as a promising autologous cell source for regenerative therapies in premature infants.