Isolation, Expansion and Characterization of Mesenchymal Stromal Cells from Fresh and Cryopreserved Umbilical Cord Tissues in Horse and Dog

Mesenchymal stromal cells (MSCs) derived from perinatal tissues represent a promising resource for regenerative medicine due to their proliferative, immunomodulatory, and differentiation properties. Among these tissues, the umbilical cord is considered an accessible and ethically acceptable source of MSCs. This study evaluated the feasibility of whole umbilical cord cryopreservation in canine and equine species and assessed the biological properties of MSCs isolated from cryopreserved tissues compared with fresh samples. Equine Wharton’s jelly and canine umbilical cord tissues were processed either immediately or after cryopreservation using two different freezing media: 10% DMSO + 10% FBS (DF) and 0.05 M glucose + 0.05 M sucrose + 1.5 M ethylene glycol (GSE). Isolated MSCs were characterized through proliferation, clonogenicity, adhesion and migration assays, trilineage differentiation, and molecular marker analysis. MSCs were successfully isolated from all experimental groups and maintained the typical morphology, plastic adherence, proliferative capacity, multilineage differentiation potential, and mesenchymal marker expression associated with MSC populations. Overall, cryopreservation did not significantly affect the main biological and functional properties of the cells. These findings support the use of cryopreserved umbilical cord tissues for veterinary biobanking and regenerative medicine applications.