An in vitro model of gonad differentiation in the chick embryo. Roller cultures in gas permeable biofoil bags.

Embryonic gonads of 6 1/2 to 12 days old chick embryos were enzymatically dissociated. The cell suspensions were cultured in small gas permeable bags of foil (Biofolie Heraeus) in a roller culture apparatus. The cells formed multiple small aggregates, in which sex specific differences developed within two days. In cell suspensions of embryonic testes smooth spheric aggregates formed with well delineated testicular cords in the center and a tunica albuginea-like mesenchymal layer at the outside. Most of the male germ cells were incorporated in the central cords. A number of germ cells were barred from entering the cords by the tunica albuginea-like mesenchymal layer and populated the outer surface of the aggregates. The aggregates of left ovary were irregular in shape and characterized by clusters of germ cells residing in an outer cortical zone. The aggregates of the right ovary, which regresses in vivo, showed poor growth and did not differentiate, thus, indicating that the suppression of right ovary was not removed in culture. In the roller cultures of dissociated embryonic gonads male and female morphogenesis was mimicked in a reproducible manner, so that the system can be used for further experimental studies of gonadal development.