Characterisation of canine CD34+/CD45 diminished cells by colony-forming unit assay and transcriptome analysis

Abstract

Haematopoietic stem and progenitor cells (HSPCs) are used for transplantation to reconstruct the haematopoietic pathways in humans receiving severe chemotherapy. However, the characteristics of canine HSPCs, such as specific surface antigens and gene expression profiles, are still unclear. This study aimed to characterise the haematopoietic ability and gene expression profiles of canine bone marrow HSPCs in healthy dogs. In this study, the CD34 positive (CD34+) cells were defined as classical HSPCs, CD34+/CD45 diminished (CD45dim) cells as more enriched HSPCs, and whole viable cells as controls. Haematopoietic abilities and gene expression profiles were evaluated using a colony-forming unit assay and RNA-sequencing analysis. Canine CD34+/CD45dim cells exhibited a significantly higher haematopoietic colony formation ability and expressed more similarity in the gene expression profiles to human and mouse HSPCs than those of the other cell fractions. Furthermore, the canine CD34+/CD45dim cells expressed candidate cell surface antigens necessary to define the canine haematopoietic hierarchy roadmap. These results indicate that the canine CD34+/CD45dim cells express the HSPC characteristics more than the other cell fractions, thereby suggesting that these cells have the potential to be used for studying haematopoietic stem cells in dogs.

In vitro aging alters the gene expression and secretome composition of canine adipose-derived mesenchymal stem cells

“…Immunophenotyping revealed the absence of CD45 protein on the surface of cAD-MSCs in all donors during the P3 stage. However, it revealed 4% of extracted cells were positive for hematopoietic stem cell marker CD34, and these cells can express the gene for PTPRC /CD45 ( 27 ). On the other hand, IL10 is a major immune regulatory cytokine with profound anti-inflammatory functions produced by various immune and non-immune cells ( 28 ).…”