A Comprehensive Protocol for Laparotomy in Swine to Facilitate Ultrasound-guided Injection into the Fetal Intraperitoneal Space

Swine are a commonly used animal model for biomedical research. One research application of swine models is the in utero injection of human or pig cells into the fetal liver (FL) or intraperitoneal space. In utero injections can be accomplished through laparotomy procedures in pregnant swine. In this study, we aimed to establish comprehensive laparotomy protocols for ultrasound-guided injections into fetuses. Two pregnant gilts, with a total of 16 fetuses, underwent laparotomy at 41 and 42 d of gestation. During surgery, we attempted to inject half of the fetuses in the FL or intraperitoneal space with saline and titanium wire for radiographic imaging after birth. After the laparotomy and fetal injections, both gilts maintained pregnancy throughout gestation and initiated labor at full term. Of the 16 fetuses present at the time of laparotomy, 12 were liveborn, 2 were stillborn, and the remaining 2 were mummies. A total of 7 fetuses from the 2 litters were known to have been injected with a wire during the surgery. After farrowing, piglets were radiographed, and 6 piglets were identified to have wire within the abdominal space. Livers were dissected, and additional radiographs were obtained. It was determined that one piglet had wire within the liver, whereas the other 5 had wire within the intraperitoneal space. Overall, we describe in-depth laparotomy surgery protocols, ultrasound-guided injection of saline and titanium wire into the FL or intraperitoneal space, postoperative monitoring protocols, and information on radiographic detection of titanium wire after piglet birth. These protocols can be followed by other research groups intending to inject cells of interest into either the intraperitoneal space or FL of fetal piglets.