The Expression Pattern of Surface Markers in Canine Adipose-Derived Mesenchymal Stem Cells

The influence of cultivation on the expression pattern of canine adipose-derived mesenchymal stem cells (cAD-MSCs) surface markers, contributing to, among others, the promotion of growth, proliferation, differentiation and immunomodulatory mechanisms of an excellent therapeutic, is still unknown. To fill the gap, we investigated CD90, CD44, CD73, CD29, CD271, CD105, CD45 and CD14 patterns of expression at the protein level with flow cytometry and mRNA level using a real-time polymerase chain reaction array. Gentle variations of expression occurred during cultivation, along with increased CD90, CD44 and CD29 expression, low and decreasing CD271 and CD73 expression and a decrease of initially high CD105. As expected, CD45 and CD14 were not expressed by cAD-MSCs. Interestingly, we discovered a significant decrease of CD73 expression, compared to early (P1-P3) to late (P4-P6) passages, although the CD73 gene expression was found to be stable. The percentage of positive cells was found to be higher for all positive markers up to P4. As CD73 ' s one important feature is a modulation from a pro-inflammatory environment to an anti-inflammatory milieu, the expression of CD73 in our conditions indicate the need to consider the time cells spend in vitro before being transplanted into patients, since it could impact their favourable therapeutical properties.

Automated summary

During cultivation, canine adipose-derived mesenchymal stem cells showed variations in the expression of surface markers, with an increase in CD90, CD44, and CD29, a decrease in CD271 and CD73, and a decrease in initially high levels of CD105. CD45 and CD14 were not expressed. Interestingly, there was a significant decrease in CD73 expression from early to late passages, despite stable gene expression. Positive marker expression was highest up to passage 4, suggesting the impact of culture time on therapeutic properties.