Mesenchymal stromal cells from equine ovarian follicular aspirates represent a promising source for cell-based therapies. However, the age of the donor mare may alter their biological properties and therapeutic potential. This study aimed to evaluate the effect of age on MSCs from ovarian follicular aspirates of young (< 10 years, n = 5) and aged (≥ 18 years, n = 5) mares. MSCs were isolated, expanded, cryopreserved at passage 3, and subsequently thawed to evaluate proliferation, immunophenotype, differentiation capacity, and gene expression profiles. Proliferation rates, total cell yield, and mean time to reach 80% confluence showed no significant differences between the age groups. Flow cytometry analysis confirmed uniform expression of MSC-associated positive markers (CD90, CD29) and absence of CD45, CD19 and MHC-II in both groups. Trilineage differentiation assays demonstrated that cells retained trilineage differentiation capacity at a qualitative level regardless of donor age. However, transcriptomic analysis using RT2 Profiler PCR Array and qPCR validation revealed substantial differences in gene expression. MSCs from young mares exhibited significant upregulation (fold change ≥ 2; p < 0.05) of stemness-related genes (LIF, POU5F2), regulators of adipogenic and chondrogenic differentiation (PPARγ, SOX9), and cell migration-associated markers (MCAM, VCAM). Conversely, MSCs from young mares showed decreased expression (fold change ≤ 0.5; p < 0.05) of lineage-specific differentiation genes (TBX5, SLC2A4), inflammatory mediators (IL6), and senescence-associated markers (ITGAX, BMP4, SMAD3). These findings suggest that, although some MSC characteristics are preserved, donor age significantly alters the transcriptomic profile of equine follicular MSCs, which could affect their therapeutic efficacy.
Keywords: Age; Follicles; Mare; Mesenchymal stromal cells; Reproduction.
Copyright © 2026 The Authors. Published by Elsevier Inc. All rights reserved.