Faculty of Land and Food Systems, University of British Columbia, Vancouver, British Columbia, Canada.
Biol Reprod. 2010 Jul;83(1):15-9. doi: 10.1095/biolreprod.110.083733. Epub 2010 Mar 17.
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.
禽类卵母细胞和胚胎的冷冻保存具有挑战性，因为卵的蛋黄结构。作为替代方法，随着有效冷冻保存方案的发展，卵巢组织冷冻保存可用于鸟类的冷冻库保存。0.5°C/min 的可编程冷冻机中以 0.5°C/min 的速度冷冻 1 周龄日本鹌鹑（ Coturnix japonica ）的卵巢组织，或者将其浸入液氮中进行玻璃化冷冻。含有慢速冷冻样本的吸管在冰水中解冻，玻璃化样本从小瓶中取出并转移到蔗糖中，在室温下依次降低浓度。通过台盼蓝测定法估计组织的细胞活力，并通过光镜检查组织组织学。从 WB（隐性羽毛颜色）雏鸡的冷冻-解冻或玻璃化-解冻组织移植到 1 周龄卵巢切除术 QO（野生型羽毛）雏鸡中，一些雏鸡接受新鲜组织作为对照组。在性成熟时，QO 接受者与 WB 雄性交配，并且 WB 后代的产生证明了冷冻保存和移植的成功。可以从通过慢速冷冻或玻璃化冷冻保存的卵巢组织中获得供体衍生的后代。本研究中使用的玻璃化方案在每个评估水平上均显示出更好的结果。这项研究表明，禽类物种的卵巢组织功能可以在亚低温下成功保存，并通过移植恢复。由于效率高且总体简单，建议使用玻璃化方案。
