Preservation of chick primordial germ cells in liquid nitrogen and subsequent production of viable offspring
Conservation of genetic material in chickens was attempted by preserving primordial germ cells in liquid nitrogen. Primordial germ cells collected from the blood of embryos at stage 13–15 of White Leghorn and Barred Plymouth Rock breeds were concentrated by Ficoll density gradient centrifugation. Th...
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Published in | Journal of reproduction & fertility Vol. 102; no. 2; pp. 321 - 325 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
England
Society for Reproduction and Fertility
01.11.1994
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Subjects | |
Online Access | Get full text |
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Summary: | Conservation of genetic material in chickens was attempted by preserving primordial germ cells in liquid nitrogen. Primordial
germ cells collected from the blood of embryos at stage 13–15 of White Leghorn and Barred Plymouth Rock breeds were concentrated
by Ficoll density gradient centrifugation. The primordial germ cells were then suspended in a freezing medium containing 10%
dimethyl sulfoxide. The temperature of the cell suspension was decreased by 1°C min −1 to −80°C; the suspension was then placed in liquid nitrogen (−196°C) and stored for 4–5 months. The cell suspension was thawed
by taking it out of liquid nitrogen and placing it in water at 4°C. The viability of the frozen–thawed primordial germ cells
was 94.2%. One hundred frozen–thawed cells were injected into the bloodstream of recipient embryos (stage 14–15) from the
other breed, from which blood had been drawn before the injection. These embryos were cultured in recipient eggshells until
hatching. Viable offspring derived from the frozen–thawed primordial germ cells were obtained by mating male and female germline
chimaeras or by mating the chimaeras with Barred Plymouth Rock chickens, and the offspring showed normal reproductive performance.
This technique for cryopreservation of primordial germ cells giving rise to viable offspring makes it possible to conserve
genetic material in avian species. |
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ISSN: | 1470-1626 0022-4251 1741-7899 |
DOI: | 10.1530/jrf.0.1020321 |