The basic study of transplantation of encapsulated cryopreserved mouse primordial follicles

我们研究了小鼠原始卵泡低温保存成功的概率。此外，我们还评估了原始卵泡包封对成功移植的有效性，并获得了以下结果。用丙二醇和蔗糖作为冷冻保护剂，用慢速冻融法成功地保存了小鼠原始卵泡。此外，经NO清除剂Hb和超氧化物清除剂SOD处理的原始卵泡在冻融后的存活率显著高于未处理组。将冷冻解冻后的卵泡包封在海藻酸盐微胶囊内，植入小鼠和大鼠腹腔。冻融后的小鼠原始卵泡移植到小鼠体内后显示出卵泡生长，但最晚期的卵泡最多有3或4层颗粒细胞。另一方面，将原始卵泡移植到大鼠体内时，未发现卵泡生长。我们证实GDF-9 mRNA在所有原始、初级和次级卵母细胞中均有表达。但本实验无法测定GDF-9 mRNA的相对量。虽然成熟卵泡和卵母细胞不能从包膜原始卵泡移植物中获得，但我们的初步结果可能提供卵巢组织和卵泡储存的重要方面。

We studied the probability of the successful cryopreservation of mouse primordial follicles. In addition, we evaluated the effectiveness of encapsulation of primordial follicles for successful transplantation, and obtained the following results.1. Mouse primordial follicles were successfully cryopreserved with slow freeze-rapid thaw protocols using propandiol and sucrose as a cryoprotectant. Furthermore, primordial follicles treated with Hb, NO scavenger, and SOD, superoxide scavenger, showed significant higher survival rates compared with non-treated group after freeze-thawing.2. Frozen-thawed follicles were encapsulated within alginate microcapsule and then implanted into peritoneal cavity in mouse and rat. Frozen-thawed mouse primordial follicules grafted in mouse demonstrated follicular growth, but the most advanced follicles had a maximum three or four granulosa cell layers. On the other hand, follicular growth was not found when primordial follicles were grafted to rats.3. We confirmed the expression of GDF-9 mRNA in all of primordial, primary and secondary oocytes. However, relative amount of GDF-9 mRNA could not determined in this experiment.Although mature follicles and oocytes were not obtained from encapsulated primordial follicle grafts, our preliminary results may offer important aspects of ovarian tissues and follicle banking.