Production of germ-line chimeras by transfer of cryopreserved gonadal primordial germ cells (gPGCs) in chicken

Abstract

Gonadal primordial germ cells (gPGCs) were collected from gonadal anlage of 5-day-old White Leghorn (WL) embryos. Collected gPGCs were suspended in freezing medium containing 10% dimethyl sulphoxide (DMSO). The cell-suspension was frozen at 1 degree C/min until the temperature reached -80 degrees C; cells were then immersed in liquid nitrogen at -196 degrees C and stored up to 3 mo. Approximately 100 frozen/thawed gPGCs were injected into the dorsal aorta of each Barred Plymouth Rock (BPR) embryo from which blood was drawn prior to germ-cell injection. The injected embryos were incubated until hatched, and hatched chicks were raised until sexually mature. Upon reaching sexual maturity, a progeny test was performed by mating recipient chicks with normal BPR of the opposite sex. Of 840 progeny produced from 4 gPGC recipients, 74 chicks (8.8%) were phenotypically white. Present results demonstrate that frozen/thawed gPGCs collected from gonadal anlage are capable of producing germ-line chimeras in chicken.