MicroRNA characterization in equine induced pluripotent stem cells.
Authors: Moro Lucia Natalia, Amin Guadalupe, Furmento Veronica, Waisman Ariel, Garate Ximena, Neiman Gabriel, La Greca Alejandro, Santín Velazque Natalia Lucia, Luzzani Carlos, Sevlever Gustavo E, Vichera Gabriel, Miriuka Santiago Gabriel
Journal: PloS one
Summary
# Editorial Summary: MicroRNA Characterisation in Equine Induced Pluripotent Stem Cells Whilst pluripotent stem cell biology is well-established in rodent and human models, equivalent knowledge in horses has remained limited—particularly regarding the microRNA regulation essential for maintaining and directing cell differentiation. Moro and colleagues successfully reprogrammed equine fibroblasts into induced pluripotent stem cells (iPSCs) using the standard Yamanaka factors (OCT-4, SOX-2, c-MYC, KLF4), then characterised two resulting cell lines (L2 and L3) through alkaline phosphatase activity, pluripotency marker expression, and immunofluorescence before examining ten key microRNAs known to regulate pluripotency in other species. The equine iPSCs demonstrated typical pluripotent behaviour—including the capacity to differentiate into all three germ layers—and exhibited significantly elevated expression of miR-302 family members alongside miR-9 and miR-96 (p<0.05), mirroring human pluripotent cells, whilst miR-145 and miR-205 were downregulated; following differentiation into embryoid bodies, miR-96 expression increased further and miR-205 reappeared in one line. Computational analysis of these microRNAs' predicted mRNA targets suggested they regulate identical biological pathways to their human equivalents—cell cycle control, epithelial–mesenchymal transition, neural specification, and pluripotency maintenance—indicating that equine stem cell regulation is fundamentally conserved. For equine professionals involved in regenerative medicine applications, these findings establish that equine iPSCs can be reliably generated and that their microRNA-mediated control mirrors human systems, providing
Read the full abstract on PubMed
Practical Takeaways
- •This foundational work enables future development of equine stem cell therapies for musculoskeletal and other tissue injuries, though clinical applications remain years away
- •Characterization of equine miRNA profiles establishes baseline knowledge for regenerative medicine applications specific to horses rather than relying on human/mouse models
- •The methodology demonstrates feasibility of generating patient-derived iPSCs from equine fibroblasts for potential personalized therapeutic approaches
Key Findings
- •Two equine iPSC lines successfully generated from fibroblasts using Yamanaka factors with confirmed pluripotency markers (OCT-4, NANOG, REX1, SOX-2, c-MYC)
- •miR-302 family, miR-9, and miR-96 showed significantly higher expression in iPSCs versus fibroblasts (p<0.05), while miR-145 and miR-205 were downregulated
- •Equine iPSCs demonstrated capacity for tri-lineage differentiation (ectodermal, endodermal, mesodermal) via embryoid bodies
- •MicroRNA target analysis suggests equine miR-302, miR-9, and miR-96 regulate similar genes as human pluripotent cells (cell cycle, neural determinants, EMT pathways)