Six- to 8-week-old male C57BL/6 mice served as recipients of donor trachea obtained from C57Bl/6 female mice of the same age and similar weight (Fig. 1 ).
Tracheas were harvested from female donors using an aseptic surgical technique. Mice were euthanized using a cocktail of ketamine (100 mg/kg), xylazine (10 mg/kg), and ketoprofen (5 mg/kg) via intraperitoneal injection. After obtaining a deep anesthetic plane, the surgical site was shaved and disinfected with alcohol and povidone iodine. A midline incision was made from the chin to the center of the chest, and subcutaneous fat pads were removed from the field. The strap muscles surrounding the trachea were separated and the trachea was dissected circumferentially. A 5-mm length of trachea was harvested and maintained in fresh phosphate-buffered saline (PBS) until implant.
Recipient male mice were prepared for transplant using the previously described surgical process for female mice. The native trachea was resected and replaced with donor female trachea using interrupted sutures of 9–0 nylon (AROS Surgical Instruments, Newport Beach, CA). After the transplanted trachea was secured in an air-tight fashion, the strap muscles were reapproximated, and the midline incision was closed using running stitches with 6–0 Vicryl suture (Ethicon, Somerville, NJ).
Postoperative care was the same as our previous study.4 Animals were evaluated daily during the first 2 weeks monitoring for weight loss >20%, stridor, and respiratory distress. Animals meeting the criteria were recommended for humane euthanasia prior to the planned endpoint and removed from the study.
Tracheas were harvested from female donors using an aseptic surgical technique. Mice were euthanized using a cocktail of ketamine (100 mg/kg), xylazine (10 mg/kg), and ketoprofen (5 mg/kg) via intraperitoneal injection. After obtaining a deep anesthetic plane, the surgical site was shaved and disinfected with alcohol and povidone iodine. A midline incision was made from the chin to the center of the chest, and subcutaneous fat pads were removed from the field. The strap muscles surrounding the trachea were separated and the trachea was dissected circumferentially. A 5-mm length of trachea was harvested and maintained in fresh phosphate-buffered saline (PBS) until implant.
Recipient male mice were prepared for transplant using the previously described surgical process for female mice. The native trachea was resected and replaced with donor female trachea using interrupted sutures of 9–0 nylon (AROS Surgical Instruments, Newport Beach, CA). After the transplanted trachea was secured in an air-tight fashion, the strap muscles were reapproximated, and the midline incision was closed using running stitches with 6–0 Vicryl suture (Ethicon, Somerville, NJ).
Postoperative care was the same as our previous study.4 Animals were evaluated daily during the first 2 weeks monitoring for weight loss >20%, stridor, and respiratory distress. Animals meeting the criteria were recommended for humane euthanasia prior to the planned endpoint and removed from the study.
