Canine Adipose Derived Mesenchymal Stem Cells Transcriptome Composition Alterations: A Step towards Standardizing...

Area of Science:

• Stem Cell Biology
• Genomics
• Veterinary Medicine

Background:

• Canine adipose-derived stem cells (cASCs) are promising for regenerative medicine.
• Previous studies focused on cASCs morphology and differentiation, but not transcriptome changes during cultivation.

Purpose of the Study:

• To characterize transcriptome alterations in undifferentiated cASCs during ex vivo cultivation (P3-P5).
• To investigate the influence of donor age on cASCs transcriptome.
• To correlate transcriptome changes with cell surface markers and differentiation potential.

Main Methods:

• Isolation and ex vivo cultivation of cASCs from six canine donors.
• Transcriptome analysis (RNA sequencing) at passages 3 (P3) and 5 (P5).
• Flow cytometry for cell surface markers (CD90, CD44).
• Osteogenic, chondrogenic, and adipogenic differentiation assays.

Main Results:

• Cultivation from P3 to P5 identified 16 differentially expressed genes.
• Co-overexpression of SOX2 and POU5F1 observed in older donors' cells.
• Upregulation of IL-6 in both young and old donor cASCs.
• Decreased CD90 and CD44 expression in higher passages (P4+).
• Successful osteo- and chondrodifferentiation, but inefficient adipodifferentiation in P3.

Conclusions:

• Ex vivo cultivation significantly impacts the transcriptome and nature of cASCs.
• Donor age influences transcriptome composition, particularly pluripotency gene expression.
• Further research correlating transcriptome with secretome is needed to evaluate therapeutic potential.