Here we describe the orthotopic rat hind-limb transplantation procedure, which seems to be the gold standard in vivo model for composite tissue allotransplantation research.
Abstract
Composite tissue allotransplantation (CTA) now represents a valid therapeutic option after the loss of a hand, forearm or digits and has become a novel therapeutic entity in reconstructive surgery. However, long term high-dose multi-drug immunosuppressive therapy is required to ensure graft survival, bearing the risk of serious side effects which halters broader application. Further progression in this field may depend on better understanding of basic immunology and ischemia reperfusion injury in composite tissue grafts.
To date, orthotopic hind limb transplantation in rats has been the preferred rodent model for reconstructive transplantation (RT), however, it is an extremely demanding procedure that requires extraordinary microsurgical skills for reattachment of vasculature, bones, muscles and nerves.
We have introduced the vascular cuff anastomosis technique to this model, providing a rapid and reliable approach to rat hind limb transplantation. This technique simplifies and shortens the surgical procedure and enables surgeons with basic microsurgical experience to successfully perform the operation with high survival and low complication rates. The technique seems to be well suited for immunological as well as ischemia reperfusion injury (IRI) studies.
Protocol
Donor Operation and Hind-Limb Harvest For the operative procedure a sterile set of microsurgical instruments is utilized. The surgeon should wear proper attire including sterile gloves, which have to be changed prior to the start of the surgical procedure. The rat is anesthetized with an intraperitoneal injection of pentobarbital (50mg/kg IP) and placed supine on a heated operation field, which facilitates a stable animal body temperature throughout the entire surgical procedure. Prio…
Discussion
Decades of experimental research have preceded and enabled successful human hand transplantation in the recent past. In 1978, Shapiro et al. developed an orthotopic hind limb transplant model in rats, well suited to study various pathological, physiological, and immunological aspects as well as functional outcome of composite tissue grafts. We have modified the most challenging part of the procedure, namely the vascular anastomosis, by introducing a non-suture cuff technique. When compared to the conventional suture tec…
Sucher, R., Oberhuber, R., Margreiter, C., Rumberg, G., Jindal, R., Lee, W. A., Margreiter, R., Pratschke, J., Schneeberger, S., Brandacher, G. Orthotopic Hind-Limb Transplantation in Rats. J. Vis. Exp. (41), e2022, doi:10.3791/2022 (2010).