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

Ji Hey Lim, Ye Eun Byeon, Hak Hyun Ryu, Yun Hyeok Jeong, Young Won Lee, Wan Hee Kim, Kyung Sun Kang, Oh Kyeong Kweon, Ji Hey Lim, Ye Eun Byeon, Hak Hyun Ryu, Yun Hyeok Jeong, Young Won Lee, Wan Hee Kim, Kyung Sun Kang, Oh Kyeong Kweon

Abstract

This study was to determine the effects of allogenic umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) and recombinant methionyl human granulocyte colony-stimulating factor (rmhGCSF) on a canine spinal cord injury model after balloon compression at the first lumbar vertebra. Twenty-five adult mongrel dogs were assigned to five groups according to treatment after a spinal cord injury: no treatment (CN); saline treatment (CP); rmhGCSF treatment (G); UCB-MSCs treatment (UCB-MSC); co-treatment (UCBG). The UCBMSCs isolated from cord blood of canine fetuses were prepared as 10(6) cells/150 microl saline. The UCB-MSCs were directly injected into the injured site of the spinal cord and rmhGCSF was administered subcutaneously 1 week after the induction of spinal cord injury. The Olby score, magnetic resonance imaging, somatosensory evoked potentials and histopathological examinations were used to evaluate the functional recovery after transplantation. The Olby scores of all groups were zero at the 0-week evaluation. At 2 week after the transplantation, the Olby scores in the groups with the UCB-MSC and UCBG were significantly higher than in the CN and CP groups. However, there were no significant differences between the UCB-MSC and UCBG groups, and between the CN and CP groups. These comparisons remained stable at 4 and 8 week after transplantation. There was significant improvement in the nerve conduction velocity based on the somatosensory evoked potentials. In addition, a distinct structural consistency of the nerve cell bodies was noted in the lesion of the spinal cord of the UCB-MSC and UCBG groups. These results suggest that transplantation of the UCB-MSCs resulted in recovery of nerve function in dogs with a spinal cord injury and may be considered as a therapeutic modality for spinal cord injury.

Figures

Fig. 1
Fig. 1
Olby scores during 8 weeks. The groups of UCB-MSC and UCBG improved more as shown with their functional scores compared with the groups of CN and CP at 2 week after transplantation (p < 0.05). The UCBG and UCB-MSC groups were significantly improved as compared with all other groups at 8 week after transplantation (p < 0.05).
Fig. 2
Fig. 2
Magnetic resonance images of the spinal cord in T2W sagittal view of the group CN (A & B), CP (C & D), G (E & F), UCB-MSC (G & H) and UCBG (I & J). Most of the dogs were shown the clear localization of the spinal cord injury lesion (circle). A, C, E, G and I: Before cell transplantation, 1 week after spinal cord injury. B, D, F, H and J: 8 weeks after cell transplantation.
Fig. 3
Fig. 3
Histopathological findings at 8 weeks after cell transplantation. (A) The epicenter of injured spinal cord of a dog in the group CP, which showed small cavity formation with a little Luxol fast blue positive area. Luxol fast blue stain. Counterstain with cresyl violet. ×12.5. (B) High magnification of A. ×40. (C) The epicenter of injured spinal cord of a dog in the group G, which was crushed and damaged in both white and grey matters with cavity formation. H&E stain. ×12.5. (D) Same lesion as C. It showed small amount of remained myelin (arrow). Luxol fast blue stain. Counterstain with cresyl violet. ×12.5. (E) The epicenter of injured spinal cord of a dog in the group UCB-MSC, which revealed abnormal structures, however it showed structural consistency with nerve cell body (circle). H&E stain. ×12.5. (F) High magnification of E (circle). H&E stain. ×400. (G) Nerve cell body in the same lesion of F (arrows). Luxol fast blue stain. Counterstain with cresyl violet. ×400.

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