Isolation and characterization of canine umbilical cord blood-derived mesenchymal stem cells

Human umbilical cord blood-derived mesenchymal stem cells (MSCs) are known to possess the potential for multiple differentiations abilities in vitro and in vivo. In canine system, studying stem cell therapy is important, but so far, stem cells from canine were not identified and characterized. In this study, we successfully isolated and characterized MSCs from the canine umbilical cord and its fetal blood. Canine MSCs (cMSCs) were grown in medium containing low glucose DMEM with 20% FBS. The cMSCs have stem cells expression patterns which are concerned with MSCs surface markers by fluorescence-activated cell sorter analysis. The cMSCs had multipotent abilities. In the neuronal differentiation study, the cMSCs expressed the neuronal markers glial fibrillary acidic protein (GFAP), neuronal class III beta tubulin (Tuj-1), neurofilament M (NF160) in the basal culture media. After neuronal differentiation, the cMSCs expressed the neuronal markers Nestin, GFAP, Tuj-1, microtubule-associated protein 2, NF160. In the osteogenic & chondrogenic differentiation studies, cMSCs were stained with alizarin red and toluidine blue staining, respectively. With osteogenic differentiation, the cMSCs presented osteoblastic differentiation genes by RT-PCR. This finding also suggests that cMSCs might have the ability to differentiate multipotentially. It was concluded that isolated MSCs from canine cord blood have multipotential differentiation abilities. Therefore, it is suggested that cMSCs may represent a be a good model system for stem cell biology and could be useful as a therapeutic modality for canine incurable or intractable diseases, including spinal cord injuries in future regenerative medicine studies.

Transplantation of canine umbilical cord blood-derived mesenchymal stem cells in experimentally induced spinal cord injured dogs

This study was to determine the effects of allogenicumbilical cord blood (UCB)-derived mesenchymal stemcells (MSCs) and recombinant methionyl humangranulocyte colony-stimulating factor (rmhGCSF) on acanine spinal cord injury model after balloon compressionat the first lumbar vertebra. Twenty-five adult mongreldogs were assigned to five groups according to treatmentafter a spinal cord injury: no treatment (CN); salinetreatment (CP); rmhGCSF treatment (G); UCB-MSCstreatment (UCB-MSC); co-treatment (UCBG). The UCB-MSCs isolated from cord blood of canine fetuses wereprepared as 10(6) cells/150microl saline. The UCB-MSCs weredirectly injected into the injured site of the spinal cord andrmhGCSF was administered subcutaneously 1 week afterthe induction of spinal cord injury. The Olby score,magnetic resonance imaging, somatosensory evokedpotentials and histopathological examinations were used toevaluate the functional recovery after transplantation. TheOlby scores of all groups were zero at the 0-week evaluation.At 2 week after the transplantation, the Olby scores in thegroups with the UCB-MSC and UCBG were significantlyhigher than in the CN and CP groups. However, there wereno significant differences between the UCB-MSC andUCBG groups, and between the CN and CP groups. Thesecomparisons remained stable at 4 and 8 week aftertransplantation. There was significant improvement in thenerve conduction velocity based on the somatosensory evokedpotentials. In addition, a distinct structural consistency ofthe nerve cell bodies was noted in the lesion of the spinalcord of the UCB-MSC and UCBG groups. These resultssuggest that transplantation of the UCB-MSCs resulted inrecovery of nerve function in dogs with a spinal cord injuryand may be considered as a therapeutic modality for spinalcord injury.