Egg Production for injections

 

To obtain a large quantitiy (>250) of eggs for injection, sexually immature FVB/N females (4-5 weeks of age) are superovulated by using consecutive PMS and HCG hormone injections. Females are mated to stud males immediately following the HCG injection.

Harvesting the eggs

 

Eggs are harvested the next day from the ampulla of the oviduct of the mated females. Eggs are treated with hyaluronidase to remove nurse cells, and are then washed through several dishes of M2 media.

Injecting the eggs

 

30-50 eggs are removed from the incubator at a time for injection. Each egg is individually injected with the DNA fragment of the day under high magnification. At the end of the injection period, eggs which have not survived injection are removed from each group.

Implanting the eggs

 

Injected eggs are then implanted in groups of 10-15 bilaterally into the oviduct of pseudopregnant females (females which have been mated to vasectomized males). They are kept under sterile conditions throughout their pregnancy.