Fundamentals of Chick Embryo Fibroblasts

This in vitro system enables to study various stages in embryonic development of chick.

It has the advantage of providing easy access to the embryo and extra-embryonic membranes for the observation of morphogenesis and growth as well as for application of different agents under study and inoculation of viruses in specific extra embryonic membranes.

The technique does not require any sophisticated equipments, media or sera and could be used not only for research purposes but also for teaching of embryology in schools, colleges and universities.

The present model may serve as an in-vitro model suitable for the fields of developmental biology, toxicology, teratology, virology and several other aspects of biomedical research.

Materials Required for Chick Embryo Fibroblasts Culture

Freshly fertilized hen’s eggs

Unfertilized hen’s eggs

Transparent glass bowl (surface diameter 8.5 cm, bottom diameter 4.5 cm and depth 4.5 cm)

Glass petri dish (9.5 cm diameter)

70% alcohol, scalpel, incubator

Procedure for Chick Embryo Fibroblasts Culture

Wash freshly fertilized hen’s eggs with water, allow to air dry and incubate at 37.5 C with a relative humidity of 70-80% for 24 h before culturing.

Allow incubated eggs to cool for 25-30 mins, wipe with 70% alcohol under laminar flow to minimize the chances of contamination from shell surface.

Keep the eggs in the horizontal position to assure that the embryo is properly positioned.

Thin albumin from unfertilized eggs was poured into a sterile bowl.

This albumin acted as a shock absorber, provided a cushion for the culture and also limited desiccation.

Crack the fertilized, incubated egg from above with the help of a scalpel at about 3-3.5 cm from the narrow end and gently release the contents of the egg over the albumin cushion in the bowl.

Transferring of the egg contents without damage to embryo and yolk and upright position of the embryo is necessary for better survival of the embryo.

Cover the bowl with a glass petridish and incubate cultures at 37.5 C and 80% humidity.

Observe the cultures after different duration of time to obtain the embryos of various Hamburger Hamilton stages.

Embryo culture can be continued for 17-19 days.

This system has been developed and used to demonstrate the glucose induced malformations in developing chick embryos.