Whole Embryo Culture of Chick, Spratt, Embryos, Alcohol, BSS, Blastoderm

www.molecular-plant-biotechnology.info

Home >> Tissue and Organ Culture >>Whole Embryo Culture of Chick

	Whole Embryo Culture of Chick - The effect of metabolic inhibitors on embryonic development was studied by Spratt (1956, 1957), using embryo cultures. In this technique, 40-hours old embryos were used and the embryo development could be followed for another 24-48 hours in vitro before the embryo dies. This involves the following steps:
(i) prepare a suitable defined medium or a synthetic medium (consult a manual), and 1ml aliquots of the medium are added to sterile watch glasses, placed on moist absorbent cotton wool pads in Petri dishes (as for organ culture); (ii) incubate hen's eggs at 38°C for 40-42 hours to provide about a dozen embryos; (iii) the shell is wiped with alcohol and broken into a sterile evaporating dish containing 50ml chick saline or BSS; (iv) a circular cut is made (using scissors) into the vitelline membrane around the blastoderm and the latter is transferred to a Petri dish containing BSS;
(v) the adherent vitelline membrane is removed with the aid of forceps and the embryo is examined under the microscope to determine the stage of development; (vi) the blastoderm is transferred to the top of the medium in the watch glass prepared in above; (vii) the blastoderm is spread on agar gel (ventral side down) and the excess BSS is removed; (viii) culture is incubated at 37.5°C.