PRODUCTION OF GERM-LINE CHIMERAS BY TRANSFER OF CRYOPRESERVED GONADALPRIMORDIAL GERM-CELLS (GPGCS) IN CHICKEN

Gonadal primordial germ cells (gPGCs) were collected from gonadal anla ge of 5-day-old White Leghorn (WL) embryos. Collected gPGCs were suspe nded in freezing medium containing 10% dimethyl sulphoxide (DMSO). The cell-suspension was frozen at 1 degrees C/min until the temperature r eached -80 degrees C; cells were then immersed in liquid nitrogen at - 196 degrees C and stored up to 3 mo. Approximately 100 frozen/thawed g PGCs 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 pr ogeny 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 t hat frozen/thawed gPGCs collected from gonadal anlage are capable of p roducing germ-line chimeras in chicken. (C) 1998 Wiley-Liss, Inc.