Abstract：
For domestic cats ovaries, recommended cold-storage limit is 24 h in Phosphate Buffered Saline (PBS) or Dulbecco`s PBS (DPBS). Here, we attempted to verify wheatear cat ovaries may benefit from more complex solutions during prolonged cold-storage (>24 h). First, the preservation capabilities of extracellular (SP+), intracellular (UW) solutions and DPBS supplemented with glutathione (DPBS+GSH) were compared using ovary fragments from the same ovary (n=10). Intact ovary stored in DPBS served as a control. Ovaries were kept at 4 °C for 48 h, and 72 h. In the second experiment, first ovary was stored in DPBS, second in SP+ or UW solution for 48 h (n = 12). Ovaries pairs stored in DPBS for 24 h served as a control (n=8). Tissue samples were evaluated directly after cold-storage and after following 24 h in vitro culture. Ovarian follicle morphology, apoptosis rates (cleaved caspase-3, TUNEL), and follicular growth activation (Ki-67) were assessed. Ovary fragmentation impaired follicular morphology preservation upon cold-storage comparing to intact ovary. However, ovarian fragments stored in UW for 48 h and in SP+ for 72 h presented better morphology than DPBS+GSH group. Comparison of intact ovaries cold-storage for 48 h showed that SP+ provided superior follicular morphology over DPBS, and it was comparable to the outcome of 24-hour storage. No follicular activation after in vitro culture was observed. Nevertheless, tissue culture increased considerably caspase-3 cleavage and TUNEL detection. The ovary fragmentation prior to cold-storage is not recommended in domestic cats. Replacement of DPBS with SP+ solution for whole ovary and UW solution for ovarian tissue fragments improves follicular structure preservation during 48-hour cold-storage.
摘要：
对于家猫卵巢，建议的冷藏限制是在磷酸盐缓冲盐水（PBS）或Dulbecco的PBS（DPBS）中24小时。这里,我们试图验证在长期冷藏（>24小时）期间，麦耳猫卵巢可能从更复杂的解决方案中受益。首先,细胞外（SP+）的保存能力，使用来自相同卵巢的卵巢片段(n=10)比较细胞内(UW)溶液和补充有谷胱甘肽的DPBS(DPBS+GSH)。储存在DPBS中的完整卵巢用作对照。卵巢在4°C下保持48小时，和72小时。在第二个实验中，第一个卵巢储存在DPBS中，在SP+或UW溶液中第二次48小时(n=12)。在DPBS中储存24小时的卵巢对用作对照(n=8)。在冷藏后和体外培养24小时后直接评估组织样品。卵泡形态,凋亡率（裂解的caspase-3，TUNEL），和卵泡生长激活（Ki-67）进行评估。与完整的卵巢相比，冷藏后的卵巢碎片损害了卵泡形态的保存。然而,卵巢碎片在UW中保存48小时，在SP中保存72小时，其形态优于DPBSGSH组。完整卵巢冷藏48小时的比较表明，SP提供优于DPBS的卵泡形态，与24小时储存的结果相当。体外培养后未观察到卵泡活化。然而，组织培养显著增加了caspase-3的裂解和TUNEL检测。在家猫中，不建议在冷藏之前进行卵巢破碎。用SP+溶液替换整个卵巢的DPBS和用于卵巢组织碎片的UW溶液改善了48小时冷藏期间的卵泡结构保存。
