The basic study of transplantation of encapsulated cryopreserved mouse primordial follicles

小鼠原始卵泡封装移植的基础研究

• 批准号: 10671510
• 负责人: SENGOKU Kazuo
• 金额: $ 1.34万
• 依托单位: ASAHIKAWA MEDICAL COLLEGE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671510/
• 关键词: Primordial follicle; Cryopreservation; Microcapsule; Oocyte maturation; 原始卵胞卵; 胚発育; HB-EGF; 活性酸素; 多精子受精防御

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.

我们研究了小鼠原始卵泡冷冻保存成功的可能性。此外，我们还评估了原始卵泡包裹对移植成功的效果，并获得了以下结果。以丙二醇和蔗糖为冷冻保护剂，采用慢速冷冻-快速解冻的方法成功地冷冻保存了小鼠原始卵泡。此外，未加清除剂HB和超氧阴离子清除剂超氧化物歧化酶处理组的原始卵泡冻融后存活率显著高于未处理组。将冷冻解冻后的卵泡包裹在海藻酸盐微囊内，然后植入小鼠和大鼠的腹膜腔内。移植到小鼠体内的冻融原始卵泡显示出卵泡生长，但最高级的卵泡最多有三到四层颗粒细胞层。另一方面，将原始卵泡移植到大鼠体内，未发现卵泡生长。我们证实GDF-9mRNA在原始卵母细胞、初级卵母细胞和次级卵母细胞中均有表达。尽管囊化原始卵泡移植不能获得成熟的卵泡和卵母细胞，但我们的初步结果可能为卵巢组织和卵泡库提供重要的方面。

项目成果

K.Sengoku: "Requirement of sperm-oocyte plasma membrane fusion for establishment of the plasma membrane block to polyspermy in human pronuclear oocytes" Molecular Reproduction Development. 52・2. 183-188 (1999)

K. Sengoku：“在人类原核卵母细胞中建立多精卵质膜阻断的精子-卵母细胞质膜融合的要求”52・2（1999）。