Production of germ-line chimeras by the transfer of cryopreserved gonadal germ cells collected from 7- and 9-day-old chick embryos |
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| Authors: | Atsushi TAJIMA Takeo MINEMATSU Mutsuo OHARA |
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| Institution: | Institute of Agriculture and Forestry, University of Tsukuba, Tsukuba-shi, Ibaraki-ken, Japan and;Hokkaido Animal Research Center, Shintoku-cho, Hokkaido, Japan |
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| Abstract: | Gonadal germ cells (GGC) were collected from the gonads of 7‐ or 9‐day‐old White Leghorn chick embryos and suspended in freezing medium containing 10% dimethylsulfoxide (DMSO). The cell suspension was frozen at ?1°C/min. until the temperature reached ?80°C. Then, the cells were immersed in liquid nitrogen at ?196°C and stored for 3–4 months. Approximately 50 frozen/thawed GGC were injected into the dorsal aorta of each 2‐day‐old Rhode Island Red (RIR) embryo, from which blood was drawn before germ‐cell injection. The injected embryos were incubated until they hatched and the chicks were raised until sexually mature. On reaching sexual maturity, a progeny test was performed by mating recipient chicks with normal RIR of the opposite sex. Progenies were obtained from male germ cell recipients that were injected with germ cells collected from 7‐ and 9‐day‐old embryos. The results demonstrated that frozen/thawed GGC collected from 7‐ or 9‐day‐old fertilized eggs can be used to produce male germ‐line chimeras. |
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| Keywords: | avian genetic resources cryopreservation germ-line chimera gonadal germ cells |
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