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Establishment of cryo-preservation method for ovary and testis and its application to storage of wild mosue genes.

卵巢和睾丸冷冻保存方法的建立及其在野生小鼠基因保存中的应用

基本信息

批准号：
06680842
负责人：
TAYA Choji
金额：
$ 0.96万
依托单位：
THE TOKYO METROPOLITAN INSTITUTE OF MEDICAL SCIENCE
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

1.Establishment of cryo-preservation method for ovary and its application to storage of wild mouse genesSuccessful freezing conditions of ovaries were detected in a series of experiments using laboratory mice. When the freezing method was applied to ovaries of MSM inbred strain derived from Japanese wild mice, mice derived from frozen ovaries were obtained, though litter sizes were very small.2.Establishment of cryo-preservation method for testis(1) Histological examinationFreezing injuries of newborn mouse testes were examined histologically at various points of a freezing course. A lot of germ cells retained a normal morphology in the program of slow cooling at a rate of 0.2 C/min till -40 C before immersion into LN_2.(2) Transplantation of frozen testisIn order to confirm the growth of germ cells and the production of spermatozoa, the frozen newborn testes were transplanted into adult testes. In the case of a freezing program described above, at 14 days after transplantation meiotic germ cells were observed and at six weeks spermatozoa were detected in the seminiferous tubules derived from the frozen testes.

1.卵巢冷冻保存方法的建立及其在野生小鼠基因保存中的应用在实验室小鼠的一系列实验中检测了卵巢成功冷冻的条件。将冷冻法应用于日本野生小鼠的MSM近交系卵巢，获得了冷冻卵巢的小鼠，但产仔量很小。睾丸冷冻保存方法的建立(1)组织学检查对新生小鼠睾丸在冷冻过程中不同时间点的冷冻损伤进行组织学检查。在以0.2 C/min缓慢冷却至-40 C的条件下，大量生殖细胞在LN_2浸泡前仍保持正常形态。(2)冷冻睾丸移植为了证实生殖细胞的生长和精子的产生，将冷冻的新生睾丸移植到成年睾丸中。在上述冷冻方案的情况下，在移植后14天观察到减数分裂生殖细胞，在6周时在冷冻睾丸的精管中检测到精子。