Non-enzymatic method of isolation of canine mesenchymal stem cells from Wharton jelly and its characterization

Wharton jelly is touted as another heavy source of the human stem cells. Considering the promising therapeutic values of stem cells in canines, this has led to the investigation of other alternative sources relevant in the extraction and identification of canine stem cells. The process of making mesenchymal stem cells (MSCs) traditionally employed in clinical apps entails collagenase-rich Wharton jelly (WJ), which has been time as well as resource-consumptive. Besides, it can lead to the presence of mechanical stress in cell isolation, and using bacterial enzymes creates a risk of contamination with endotoxins and xenoantigens. Thus, during the current research, we chose explant culture method as a more efficient and easier yet effective method of isolating MSCs in canine Wharton jelly. The cells were isolated and adherent to plastic and they had a fibroblastoid shape. Larger studies which have evaluated the adult mesenchymal stem cells in vitro conditions that allow trilineage differentiation show homogeneous trend of the gene-expression profiles they are analysing. The results of the present study help to determine that this patient demonstrates positive phenotype condition (manifestation of CD44, CD73, non-ticketed CD90 and incapability of presenting edging CD34). All these data help in the definition of mesenchymal stem cells. This was further confirmed by immunofluorescence labelling, which pointed at strong expression of MSC specific markers. Our findings show that explant culture-method allows the easy isolation of canine WJ-MSCs without affecting their morphology and multipotensoriness.