Comparative Transcriptome and MicroRNA Profiles of Equine Mesenchymal Stem Cells, Fibroblasts, and Their...

Area of Science:

• Veterinary Medicine
• Cell Biology
• Regenerative Medicine

Background:

• Mesenchymal stem cells (MSCs) are crucial for regenerative medicine due to their paracrine signaling.
• Extracellular vesicles (EVs) from MSCs carry bioactive molecules, mediating intercellular communication.
• Understanding MSC and EV profiles is key to optimizing equine regenerative therapies.

Purpose of the Study:

• To compare the transcriptomic (mRNA and miRNA) profiles of equine adipose tissue-derived MSCs (AT-MSCs) and bone marrow-derived MSCs (BM-MSCs).
• To characterize the miRNA content of EVs secreted by equine AT-MSCs, BM-MSCs, and ovarian fibroblasts.
• To investigate how MSC tissue origin influences their transcriptomic profiles and EV cargo.

Main Methods:

• Equine AT-MSCs, BM-MSCs, and ovarian fibroblasts were characterized using ISCT criteria.
• Extracellular vesicles (EVs) were isolated via ultracentrifugation and validated using nanoparticle tracking analysis and flow cytometry.
• Next-generation sequencing was employed to analyze mRNA and miRNA profiles of cells and their secreted EVs.

Main Results:

• All equine cell types met MSC criteria, exhibiting distinct mRNA and miRNA profiles.
• EVs contained diverse RNA species, including novel miRNAs unique to AT-MSCs and BM-MSCs.
• Significant differences in miRNA expression were observed between MSC types and fibroblasts, correlating with tissue origin.

Conclusions:

• Equine MSC tissue origin significantly impacts cellular transcriptomic profiles and the miRNA composition of secreted EVs.
• These origin-specific molecular signatures are crucial for understanding and predicting the therapeutic potential of equine MSCs.
• Identifying key molecular markers can guide the optimization of MSC-based regenerative therapies in horses.