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.

我们研究了微珠法保存未成熟卵泡的可能性。将冷冻保存的小鼠原始细胞包裹在琼脂糖凝胶或三层微球中，然后将微球异种植入大鼠体内。取得了以下研究成果：1.用慢速冷冻-快速解冻方法成功地冷冻保存了小鼠原始卵泡。此外，添加自由基清除剂，特别是在冷冻和解冻介质中添加不含自由基清除剂的超氧化物歧化酶，可以提高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球在同种异体移植后保持了生物相容性，提示该方法可能在未来应用于保存未成熟卵泡。但异种移植后，未获得成活的卵泡和卵泡生长。GDF9和C-kit基因在未成熟卵母细胞中的表达已得到证实。尽管微球技术在冷冻保存的小鼠未成熟卵泡异种移植中未获得成功，但我们的初步结果可能为该微珠法在卵巢组织库中的应用提供重要的方面。

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。