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The objective of this study was to graft autologous mesenchymal stem cells (MSCs) at the site of surgical repair of a soft palate defect in an adult horse in an attempt to improve wound healing and to investigate whether the transplanted MSCs would integrate into the soft palate structure and participate in regeneration. Bone marrow was collected from an adult horse with a full-thickness soft palate defect. The MSCs were isolated, cultured in monolayers, and labeled with 5-bromo-2-desoxymidine (BrdU) and chloromethylbenzamido-DiI-derived (cm-DiI) before transplantation. The soft palate defect was repaired by mandibular symphysiotomy, and the labeled MSCs were injected into the repaired soft palate. Postmortem examination revealed that 90% of the soft palate defect had been sutured. Staining by BrdU and cm-DiI was intense in the soft palate tissue. Labeled MSCs were detected in tissue slices from the injection sites. The cells were organized in a manner similar to that in native soft palate tissue, indicating successful engraftment.