This article describes the methodology for retrieving perinatal mouse embryos, which serve as model systems for studying early mammalian embryogenesis. The process involves careful dissection and handling of the embryos to ensure their viability for research purposes.
Mouse embryos are used as model systems for studying early mammalian embryogenesis. To retrieve perinatal mouse embryos at the desired stage of embryonic development, begin with a euthanized pregnant mouse in the supine position.
Next, sterilize its abdominal area and make a transverse incision on the abdominal wall to reveal the underlying uterine horns. The uterine horns are long tubular structures containing a string of embryos. Now, carefully separate the uterine horns from the abdomen by excising them from the surrounding tissue.
Transfer the harvested uterine horns into a dish containing buffer. Rinse the uterine horns to remove tissue debris and blood. Further, incise between the implantation sites along the uterine horns to separate each embryo. Excise the decidua, the maternal placental layer, along with the embryo-derived placental disc.
Next, remove the small extraembryonic membranous structure surrounding the embryo that provides nutrition to it - the yolk sac. Finally, remove the amniotic membrane - the innermost extraembryonic membrane consisting of a fluid-filled sac that surrounds and protects the embryo - to release the mouse embryo.
The recovered perinatal embryos can be used for studying early mammalian development.
Clean dissecting scissors and forceps with 70% ethanol. To collect perinatal embryos at E18.5 or E19.5, first, euthanize pregnant mice using carbon dioxide inhalation. Then, perform cervical dislocation according to NIH guidelines.
Next, spray 70% ethanol on the ventral abdominal surface of the animal. Then, using the sterile dissecting scissors and forceps, open the abdominal cavity eventually. Lift the entire uterus and separate it from the body at the tips of the uterine horns. Then, rinse the entire uterus with 1x PBS in a Petri dish.
Cut the uterus segmentally with dissecting scissors and remove the placental decidua with dissecting forceps to expose the embryos in yolk sacs. Remove the yolk sac first and then remove the amniotic membrane with dissecting forceps to liberate the perinatal embryos.
繁體中文
