title: Long-term culture of chicken primordial germ cells isolated from embryonic blood and production of germline chimaeric chickens.
abstract: Production of germline chimaeric chickens by the transfer of cultured primordial germ cells (PGC) is a useful system for germline manipulation. A novel culture system was developed for chicken PGC isolated from embryonic blood. The isolated PGC were cultured on feeder cells derived from chicken embryonic fibroblast. The cultured PGC formed colonies and they proliferated about 300-times during the first 30 days. The cultured PGC retained the ability to migrate to recipient gonads and were also chicken VASA homologue (CVH)-positive. Female PGC were present in the mixed-sex PGC populations cultured for more than 90 days and gave rise to viable offspring efficiently via germline chimaeric chickens. Male cultured PGC were transferred to recipient embryos and produced putative chimaeric chickens. The DNA derived from the cultured PGC was detected in the sperm samples of male putative chimaeric chickens, but no donor derived offspring were obtained. Donor-derived offspring were also obtained from germline chimaeric chickens by the transfer of frozen-thawed cultured PGC. The culture method for PGC developed in the present study is useful for manipulation of the germline in chickens, such as preservation of genetic resources and gene transfer.
authors.affiliation: National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan. Electronic address: mnaito@affrc.go.jp.
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Naito
authors.affiliation: National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Harumi
authors.affiliation: International Institute of Avian Conservation Science, P.O. Box 47087, Abu Dhabi, United Arab Emirates.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kuwana
mesh_terms: D000818:Animals; D001773:Blood Cells / Q000166:cytology*; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D002642:Chick Embryo / Q000166:cytology*; D002645:Chickens; D002678:Chimera; D005260:Female; D005854:Germ Cells / Q000166:cytology*; D008297:Male; D061251:Primary Cell Culture / Q000379:methods* / Q000662:veterinary
keywords: Chicken; Culture; Embryo; Germline chimaera; Primordial germ cell