Production of Donor-Derived Offspring from Cryopreserved Ovarian Tissue in Japanese Quail (Coturnix japonica)1

Abstract

Cryopreservation of avian ova and embryos is challenging because of the yolky structure of the egg. As an alternative, with the development of effective cryopreservation protocols, ovarian tissue cryopreservation could be used for cryobanking for birds. Pieces of ovarian tissue of week-old Japanese quail (Coturnix japonica) were frozen at 0.5 degrees C/min in a programmable freezer or vitrified by immersion in liquid nitrogen. Straws containing slow-frozen samples were thawed in ice water, and vitrified samples were removed from the vials and transferred into sucrose, with the concentration lowered in sequence at room temperature. Cell viability of tissue was estimated by trypan blue assay, and tissue histology was examined by light microscopy. Frozen-thawed or vitrified-warmed tissue from WB (recessive plumage color) chicks was transplanted into week-old ovariectomized QO (wild-type plumage) chicks, with some chicks receiving fresh tissue as a control group. At sexual maturity, QO recipients were mated to WB males, and the production of WB offspring demonstrated successful cryopreservation and transplantation. Donor-derived offspring were obtained from the ovarian tissue that had been cryopreserved by either slow-freezing or vitrification. The vitrification protocol used in this study showed better outcomes at each level of evaluation. This study demonstrated that the function of ovarian tissue in avian species can be successfully preserved at subzero temperatures and recovered by transplantation. The vitrification protocol is recommended because of high efficiency and overall simplicity.

Production of Live Offspring from Testicular Tissue Cryopreserved by Vitrification Procedures in Japanese Quail (Coturnix japonica)1

“…Using vitrification for preservation of testicular tissue has been attempted in mammalian species [9,10], including humans [11] and fish [12], with encouraging results, although no offspring have been produced. The effectiveness of vitrification procedures in cryopreserving ovarian tissue of Japanese quail has been demonstrated [13]. Testicular tissue of Japanese quail has also been cryopreserved using vitrification procedures and shows normal macro-and microscopic structures after shortterm in ovo culture [14].…”

Comparative costs of programmes to conserve chicken genetic variation based on maintaining living populations or storing cryopreserved material

“…Economies of scale can be expected for larger numbers of lines. Liu et al (2010) vitrified and warmed quail ovarian tissue and five of the seven recipients that survived surgery produced donor-derived offspring. Song and Silversides (2007a) used fresh tissue to generate donor-derived offspring from three of nine chickens operated on.…”

“…Ovaries can be recovered by transplantation (Song and Silversides, 2007a;Liu et al, 2010) and the hosts can be inseminated using cryopreserved semen. The entire process of recovery takes less than a year, suggesting that stored germplasm can be accessed and used to generate a population that is used during the time-frame of a short-term grant, allowing the cost of recovery to be included in the budget.…”

“…Recently, techniques for cryopreserving and transplanting mammalian gonads (Gunasena et al, 1997;Honaramooz et al, 2002;Schlatt et al, 2002) have been adapted for use with chickens (Song and Silversides, 2007a, b, c) and Japanese quail (Song and Silversides, 2008;Liu et al, 2010). Liu et al (2010) produced donorderived quail chicks from 50% of the hens receiving vitrified ovarian tissue, which suggests that tissue transplantation can be utilised effectively in the conservation of avian genetic resources.…”

“…Liu et al (2010) produced donorderived quail chicks from 50% of the hens receiving vitrified ovarian tissue, which suggests that tissue transplantation can be utilised effectively in the conservation of avian genetic resources.…”