Prenatal Mouse Embryos Retrieval: A Procedure to Harvest Embryos from Pregnant Mouse

Abstract

Source: Yun, K. Assessing Urinary Tract Junction Obstruction Defects by Methylene Blue Dye Injection. J. Vis. Exp. (2017)

This video demonstrates the retrieval of embryos from a pregnant mouse. The recovered perinatal embryos can be used as model systems for studying mammalian development.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Dissection of Prenatal Embryos

1. Clean dissecting scissors and forceps with 70% ethanol.
2. To collect perinatal embryos at E18.5 or E19.5, euthanize pregnant mice first using CO₂ inhalation and then perform cervical dislocation according to ethical guidelines.
   NOTE: The pregnant female mouse usually gives birth starting at E18.5, however, larger kidneys are easier to manipulate. Therefore, kidneys at E19.5 closer to birth are easier to perform this analysis on. However, pups with bilateral urinary tract obstructions die after birth. Depending on the characteristics of experimental mice, an appropriate collection day/time should be empirically determined.
3. Spray 70% ethanol on the ventral abdominal surface and then open the abdominal cavity ventrally using dissecting scissors and forceps.
4. Lift the entire uterus and separate it from the body by cutting with dissecting scissors at the tips of the uterine horns.
5. Rinse the entire uterus with 1x PBS in a petri dish.
6. Cut the uterus segmentally with dissecting scissors and remove placental decidua with dissecting forceps to expose the embryos in yolk sacs.
7. Remove the yolk sac first and then remove the amniotic membrane with dissecting forceps to liberate the perinatal embryos.

Materials

Delicate Operating Scissors	Roboz Surgical Instrument Co.	RS-6702
Micro Dissecting Forceps	Roboz Surgical Instrument Co.	RS-5135
Dumont Tweezers; Pattern #5	Roboz Surgical Instrument Co.	RS-5045
Quality Biological Inc. NORMAL SALINE – 500ML	Fisherscientific	50-983-204