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.

研究了小鼠原始卵泡冷冻保存成功的可能性。此外，我们还对原始卵泡包裹技术在移植成功中的有效性进行了评价，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-快速解冻法成功地冷冻保存了小鼠原始卵泡。此外，经血红蛋白、一氧化氮清除剂和超氧化物歧化酶处理的原始卵泡冻融后的存活率显著高于未处理组。将冻融后的卵泡包裹在海藻酸钠微囊内，然后植入小鼠和大鼠腹腔。小鼠移植的原始卵泡经冻融后仍有卵泡生长，但最高级的卵泡最多有三层或四层颗粒细胞。另一方面，原始卵泡移植后未见卵泡生长.我们证实了GDF-9 mRNA在原始卵母细胞、初级卵母细胞和次级卵母细胞中的表达。尽管未从包裹的原始卵泡移植物中获得成熟卵泡和卵母细胞，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）。