ABSTRACT.Regenerative medicine using bone marrow cells is an attractive therapy for the cure of patients with severe liver disease. Here, we show the therapeutic potential of canine bone marrow stromal cells (BMSCs) in mouse models of CCl 4 -induced chronic liver dysfunction. We used two different models for xenotransplantation, nude mice and cyclosporine A (CSA) immunosuppressed mice. Serum parameters from a standard liver panel were not improved following transplantation. However, fibrotic liver lesions with severe inflammation were decreased in CCl 4 -treated CSA mice following BMSC transplantation. Effective migration of transplanted canine BMSCs was limited to persistently injured liver in CCl 4 -treated CSA mice, where they may be effective in resolving inflammatory fibrotic lesions. These results suggest that canine BMSCs are an effective cell source for liver regeneration.

Regenerative therapy using bone marrow stromal cells (BMSCs) has begun to be clinically applied in humans and dogs for neurological disorders such as spinal cord injury. Under appropriate conditions in vitro, BMSCs differentiate into neuronal cells, which may improve the effects of regenerative therapy. In this study, we evaluated canine neuron-like cells (NLCs) derived from BMSCs. We speculated on their suitability for neuro-transplantation from the point of view of their morphological features, long-term viability, abundant availability, and ability to be subcultured. Canine NLCs were differentiated as follows: third-passage BMSCs were maintained in pre-induction medium containing 2-mercaptoethanol and dimethylsulfoxide for 5 h, and then cells were transferred to neuronal induction medium containing fetal bovine serum, basic fibroblast growth factor, epidermal growth factor, dibutyryl cyclic AMP, and isobutylmethylxanthine for 7 or 14 days. Canine NLCs fulfilled the transplantation criteria and expressed markers of both immature neurons (nestin, 84.7 %) and mature neuronal cells (microtubule-associated protein-2, 95.7 %; βIII-tubulin protein, 12.9 %; glial fibrillary acidic protein, 9.2 %). These results suggest that canine BMSCs can be induced to differentiate into neuronal cells and may be suitable for neuro-transplantation. This study may provide information for improving cellular therapy for neurological diseases.

Bone marrow stromal cell (BMSC) transplantation has been reported as treatments that promote functional recovery after spinal cord injury (SCI) in humans and animals. Polyethylene glycol (PEG) has been also reported as treatments that promote functional recovery after spinal cord injury (SCI) in humans and animals. Therefore, administration of PEG combined with BMSC transplantation may improve outcomes compared with BMSC transplantation only in SCI model mice. SCI mice were divided into a control-group, BMSC-group, PEG-group and BMSC+PEG-group. BMSC transplantation and PEG administration were performed immediately after surgery. Compared to the control-group, PEG- and BMSC+PEG-groups showed significant locomotor functional recovery 4 weeks after therapy. We observed no significant differences among the groups. In the BMSC- and BMSC+PEG-groups, immunohistochemistry showed that many neuronal cells aggressively migrated toward the glial scar from the region rostral of the lesion site. In the control- and PEG-groups, the boundary of the injured regions was covered with astrocytes, and a few neuronal cells were migrated toward the glial scar. We conclude that combined BMSC transplantation with PEG treatment showed no synergistic effects on locomotor functional recovery or beneficial cellular events. Further studies may improve the effect of the treatment, including modification of the timing of BMSC transplantation.

Regenerative therapy has begun to be clinically applied in humans and dogs to treat neurological disorders, such as spinal cord injury (SCI). Here, we show the therapeutic potential of transplantation of cultured canine bone marrow stromal cells (BMSCs) into mice with SCI. Canine BMSC transplantation therapy was performed, immediately after the spinal cord was injured. Canine BMSC therapy enhanced functional recovery of the hind limbs in mice with SCI. Nestin-positive cells were observed only in the lesion of mice with SCI that received BMSCs. These results suggest that canine BMSCs promote functional recovery in mice with SCI and that migration of nestin-positive cells may contribute to the efficacy of the BMSC treatment.

ABSTRACT. Autologous bone marrow stromal cells (BMSCs) infusion therapy improves the hepatic fibrosis. To investigate the mechanism of remission, we evaluated the matrix metalloproteinase (MMP)-2 and -9 activity in canine BMSCs and the effect of pro-inflammatory cytokines on their expression. The activity and the gene expression of MMPs were analyzed by gelatin zymography and quantitative RT-PCR, respectively. The specific gelatinase bands were indicative effect of MMP-2 and -9 in canine BMSCs. MMP-2 expression seemed to be increased by TNF-α and IL-1β while MMP-9 was enhanced by TNF-α and IL-6. These results suggested that remissive effect on liver fibrosis might be partly attributable to the MMP-2 and -9 activity in BMSCs under the inflammatory condition. KEY WORDS: bone marrow stromal cell (BMSC), canine, culture, inflammatory cytokines, matrix metalloproteinase (MMPs).

A thirteen-month-old French bulldog was referred for evaluation of a protrusion in the pubic region. Physical examination, radiographs and histopathological testing of both gonads following a gonadectomy indicated that the dog was a male pseudohermaphrodite. The karyotype was 78, XX chromosome, and the canine SRY gene was negative. Therefore, the present case was diagnosed as a SRY negative XX male in a French bulldog.

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