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Method for improving vitrification freezing efficiency of ovarian tissues

A technology for vitrification and freezing of ovarian tissue, which can be used in biochemical equipment and methods, tissue culture, preservation of human or animal bodies, etc., and can solve problems such as low pregnancy rate

Active Publication Date: 2020-10-30
ZHEJIANG UNIV
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Problems solved by technology

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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  • Method for improving vitrification freezing efficiency of ovarian tissues
  • Method for improving vitrification freezing efficiency of ovarian tissues
  • Method for improving vitrification freezing efficiency of ovarian tissues

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Embodiment Construction

[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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Abstract

The invention relates to a fertility protection and preservation technology. The invention discloses a method for improving vitrification freezing efficiency of ovarian tissues. The method comprises the following steps: 1) freezing the ovarian tissues: melatonin is added into a basic solution, an equilibrium solution and a freezing solution respectively, and the concentration of the melatonin reaches 0.01 mM; then, the ovarian tissues taken from a human body are immersed in the basic solution, the equilibrium solution and the freezing solution respectively for 5-20 min in sequence, and then the ovarian tissues are put into freezing tubes respectively to be sealed and directly put into liquid nitrogen to be frozen and stored; and (2) unfreezing the ovarian tissues, namely respectively adding melatonin into a 1mol / L sucrose solution, a 0.5 mol / L sucrose solution and a 0.25 mol / L sucrose solution to obtain three resuscitation solutions with the melatonin concentration of 0.01 mM, and preheating at 37 DEG C. The method has the characteristics that apoptosis of granular cells in the unfrozen ovarian tissues can be reduced, and the vitrification freezing efficiency of the ovarian tissuesis improved.

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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Application Information

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Patent Type & Authority Applications(China)
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
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