Abe Yasuyuki, Asano Tomoyoshi, Ali Mohammed, Suzuki Hiroshi
Obihiro University of Agriculture and Veterinary Medicine 2-13 Inada-cho 080-8555 Obihiro Hokkaido Japan.
Reprod Med Biol. 2010 Feb 2;9(2):115-120. doi: 10.1007/s12522-010-0045-6. eCollection 2010 Jun.
The effects of the cryoprotectant and the container (holder) used for the vitrification of canine germinal vesicle stage oocytes were examined to improve the cryopreservation method for canine oocytes and embryos.
Canine cumulus-oocyte complexes (COCs) were collected from ovaries, and were vitrified with E30S (30% ethylene glycol and 0.5 M sucrose) or DAP213 (2 M dimethyl sulfoxide, 1 M acetamide, and 3 M propylene glycol) solution held by a cryotube or cryotop sheets. After warming, the oocytes were stained with propidium iodide for the assessment of their plasma membrane integrity.
In all the vitrification groups, more than 65% of the vitrified oocytes displayed a normal morphology (E30S-top, 65.6%; DAP-tube, 67.3%; DAP-top, 80.0%). However, when assessed by propidium iodide staining, the viability of oocytes in the DAP-top group (43.6%) was higher than that in the E30S-top group (21.3%, < 0.05). Furthermore, the viability of the oocytes in the DAP-top group (43.6%) was higher than that in the DAP-tube group (4.1%, < 0.05).
These results suggest that a combination of DAP213 as the cryoprotectant and a cryotop sheet as the holder improved viability after the vitrification of canine oocytes at the germinal vesicle stage.
研究冷冻保护剂和用于犬原始卵泡期卵母细胞玻璃化的容器（支架）的效果，以改进犬卵母细胞和胚胎的冷冻保存方法。
从卵巢中采集犬卵丘-卵母细胞复合体（COCs），并用E30S（30%乙二醇和0.5M蔗糖）或DAP213（2M二甲基亚砜、1M乙酰胺和3M丙二醇）溶液在冷冻管或冷冻载片上进行玻璃化。解冻后，用碘化丙啶对卵母细胞进行染色，以评估其质膜完整性。
在所有玻璃化组中，超过65%的玻璃化卵母细胞形态正常（E30S-载片组，65.6%；DAP-冷冻管组，67.3%；DAP-载片组，80.0%）。然而，通过碘化丙啶染色评估时，DAP-载片组卵母细胞的存活率（43.6%）高于E30S-载片组（21.3%，P<0.05）。此外，DAP-载片组卵母细胞的存活率（43.6%）高于DAP-冷冻管组（4.1%，P<0.05）。
这些结果表明，DAP213作为冷冻保护剂与冷冻载片作为支架相结合，可提高犬原始卵泡期卵母细胞玻璃化后的存活率。
