A method for improving the efficiency of vitrification of ovarian tissue
A technology of vitrification and ovarian tissue, applied in biochemical equipment and methods, tissue culture, preservation of human or animal bodies, etc., can solve the problem of low pregnancy rate, achieve short time-consuming, simple method, and reduce apoptosis Effect
Active Publication Date: 2021-09-24
ZHEJIANG UNIV
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As of June 2017, the number of live births reported in the world has exceeded 130, and it has increased logarithmically from 2016 to 2017. It can be seen that the clinical application of ovarian cryotransplantation technology is feasible, but there is still a low pregnancy rate after transplantation. question
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[0046]The applicant found that adding melatonin to both the vitrification solution and the resuscitation solution can protect vitrified follicles and granulosa cells from apoptosis in ovarian tissue. Therefore, the present invention intends to evaluate its effectiveness by adding 0.01 mM melatonin during vitrification of mouse ovary tissue and adding 0.01 mM melatonin during recovery, compared with conventional vitrification methods.
[0047] The present invention has adopted mouse ovary tissue to carry out the experiment of vitrification and recovery; Specifically as follows:
[0048] 1. Obtaining mouse ovary tissue: Collect ovaries of 6-8 week old female ICR mice, kill by neck dislocation, remove both ovaries and put them in sterile PBS immediately. Rinse repeatedly in a Petri dish with PBS to remove blood from the tissue, and remove fat and connective tissue around the ovary.
[0049] 2. Freezing of mouse ovary tissue: Vitrification method was used to treat the ovarian tis...
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The invention relates to a fertility protection preservation technology. The technical solution is: a method for improving the vitrification efficiency of ovarian tissue; it is carried out according to the following steps: 1) Ovarian tissue freezing: firstly add melatonin to the base liquid, the balance liquid, and the freezing liquid respectively, so that the concentration of the melatonin reaches 0.01mM; then immerse the ovarian tissue taken from the human body in the base solution, balance solution, and freezing solution for 5-20 minutes, and then put them into cryovials, seal them, and put them directly into liquid nitrogen for cryopreservation; 2) Ovary Tissue thawing: first add melatonin to 1mol / L sucrose solution, 0.5mol / L sucrose solution and 0.25mol / L sucrose solution to obtain three resuscitation solutions with melatonin concentration of 0.01mM, and then Preheat to 37°C. The method should have the characteristics of reducing the apoptosis of granulosa cells in ovarian tissue after thawing and improving the efficiency of vitrification of ovarian tissue.
Description
technical field [0001] The invention relates to a fertility protection preservation technology, in particular to a method for improving vitrification efficiency of ovarian tissue. Background technique [0002] As the incidence of malignant tumors increases year by year, the age of onset is gradually younger, and the age of childbearing is delayed due to personal reasons, the demand for fertility preservation has increased dramatically. Methods to maintain female fertility mainly include embryo freezing, egg freezing and ovarian tissue freezing. Ovarian cryopreservation has its unique advantages in fertility preservation, for example, it can preserve the reproductive function of prepubertal female patients, can be performed at any time without delaying the treatment of the tumor itself, and can preserve the reproductive function of patients with hormone-sensitive tumors such as breast cancer. Maintain female gonadal endocrine function, etc. In 2004, Donnez et al. reported t...
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IPC IPC(8): A01N1/02C12N5/071
CPCA01N1/021A01N1/0221A01N1/0226C12N5/0682C12N2500/34C12N2501/30
Inventor 张松英施丽冰危先江竺海燕
Owner ZHEJIANG UNIV