The basic study of xenotransplantation of encapsulated cryopreserved mouse immature follicles

封装冻存小鼠未成熟卵泡异种移植的基础研究

• 批准号: 12671574
• 负责人: SENGOKU Kazuo
• 金额: $ 2.05万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671574/
• 关键词: Primordial follicle; Microcapsule; cryopreservation; Oocyte factor; free radical; xenotransplantation

We studied the possibility of preservation of immature ovarian follicles using microbeads methods. The cryopreserved mouse primordial was encapsulated in agarose or three-layer microbeads, and microbeads were xenogenically implanted in the rats. The following results were obtained.1. Mouse primordial follicles were successfully cryopreserved with slow freeze-rapid thaw protocols. Moreover, the supplementation of free radical scavengers, particularly combinations of SOD with NO scavengers in freezing and thawing media, improved the post-thaw survival and fertilization rates of cryopreserved mouse oocytes.2. The microbeads encapsulating immature follicles retain biocompatibility after homotransplantations, suggesting possible future application of this methods for preservation immature follicles. However, after xenotransplantation, viable follicles and follicular growth were not obtained.3. The expression of mRNA of GDF9 and C-kit in the immature oocytes was confirmed.Although successful xenotransplantation was not obtained using encapsulated cryopreserved mouse immature follicles, our preliminary results may offer important aspects of future applications of this microbeads method for ovarian tissues banking.

我们研究了用微珠法保存未成熟卵巢卵泡的可能性。用琼脂糖或三层微球包封小鼠原始细胞，异种植入大鼠体内。得到了以下结果：采用慢速冻融技术成功保存小鼠原始卵泡。此外，在冷冻和解冻培养基中添加自由基清除剂，特别是SOD与NO清除剂的组合，可以提高冷冻保存的小鼠卵母细胞的解冻后存活率和受精率。包裹未成熟卵泡的微珠在同型移植后仍能保持生物相容性，这表明这种方法可能在未来用于保存未成熟卵泡。然而，异种移植后，未获得可存活的卵泡和卵泡生长。证实了GDF9和C-kit mRNA在未成熟卵母细胞中的表达。尽管使用冷冻保存的小鼠未成熟卵泡没有获得成功的异种移植，但我们的初步结果可能为这种微珠方法在卵巢组织库中的未来应用提供了重要的方面。

项目成果

Sengoku K et al.: "Requirement of nitric oxide for murine oocyte maturation, embryo development, and trophoblast outgrowth in vitro"Molecular Reproduction and Development. 58(3). 262-268 (2001)

Sengoku K 等人：“小鼠卵母细胞成熟、胚胎发育和体外滋养层生长所需的一氧化氮”分子生殖和发育。

S.Dinara, K.Segoku et al.: "Effects of supplementation with free radical scavengers on the survival and fertilization rates of mouse cryopreserved oocytes"Human Reproduction. 16/9. 1976-1981 (2001)

S.Dinara、K.Segoku 等人：“补充自由基清除剂对小鼠冷冻卵母细胞的存活率和受精率的影响”人类生殖。

Sengoku K et al.: "Requirement of nitric oxide for murine oocyte maturation, embryo development, and trophoblast outgrowth in vitro."Mol Reprod Dev. 58(3). 262-268 (2001)

Sengoku K 等人：“小鼠卵母细胞成熟、胚胎发育和体外滋养层生长所需的一氧化氮。”Mol Reprod Dev。

S.Dinara, K.Sengoku et al.: "Effects of supplementation withnfree radical scavengers on the survival and fertilization rates of mouse cryopreserved oocytes"Human Reproduction. 16(9). 1976-1981 (2001)

S.Dinara、K.Sengoku 等人：“补充自由基清除剂对小鼠冷冻卵母细胞的存活率和受精率的影响”人类生殖。

Sengoku K, et al.: "Requirement of nitric oxide for murine oocyte maturation, embryo development, and trophoblast outgrowth in vitro"Molecular Reproduction and Development. 58(3). 262-268 (2001)

Sengoku K 等人：“小鼠卵母细胞成熟、胚胎发育和体外滋养层生长所需的一氧化氮”分子生殖和发育。