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Programmed cryopreservation method for ovarian tissues

A technology for ovarian tissue and cryoprotectant, which is applied in the field of programmed cryopreservation of mammalian ovarian tissue, and can solve problems such as optimization problems of programmed freezing scheme

Inactive Publication Date: 2021-02-02
符晓倩
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although programmed freezing has achieved good results so far, there are many challenges in the optimization of programmed freezing protocols

Method used

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  • Programmed cryopreservation method for ovarian tissues
  • Programmed cryopreservation method for ovarian tissues

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experiment example

[0028] 1. Preparation of ovarian tissue freezing solution, thawing solution and culture medium

[0029] The solutions used for freezing and thawing ovarian tissue are prepared by adding different kinds of cryoprotectants with different concentrations to the freeze-thaw base solution. All solutions were sterilized by filtration through 0.22 μm pore size filters.

[0030] 1.1 Freeze-thaw basal fluid: HEPES-buffered human tubal fluid (human tubal fluid, HTF) containing 5mg / ml human serum albumin (human serum albumin, HSA): add 5ml HAS to 95ml HEPES-HTF culture medium.

[0031] 1.2 Thawing resuscitation solution: add 3.4g sucrose to 10ml freeze-thaw base solution, the final concentration of sucrose is 1M.

[0032] 1.3 The first cryoprotectant solution: 110 μl of PROH was added to 890 μl of freeze-thawed base solution, and the final concentration of PROH was 1.5M.

[0033] 1.4 The second cryoprotectant solution: Add 110 μl of PROH and 100 μl of thawed resuscitation solution to 79...

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Abstract

The invention relates to a technical system for programmed cryopreservation of mammal ovarian tissue, and belongs to the technical field of tissue cryopreservation. The ovarian tissue frozen and resuscitated by the method can keep good physiological activity. Ovarian tissue freezing not only can preserve the function of ovarian generation gametes, but also can keep the endocrine function of the ovarian generation gametes, and has incomparable advantages in fertility preservation. After the ovarian tissue is obtained and treated, the ovarian tissue is balanced by a cryoprotectant and cooled bya programmed freezing instrument, and the ovarian tissue is stored in liquid nitrogen. By improving the components and proportion of the cryoprotectant, the balance time and the freezing and cooling procedures, follicles in ovarian tissues can be well protected from being damaged by freezing resuscitation stimulation, and a feasible method is provided for protecting and storing the fertility of mammals.

Description

technical field [0001] The invention relates to a technical system for programmed cryopreservation of mammalian ovarian tissue, which can maintain good physiological activity of mammalian ovarian tissue after freezing and thawing, and belongs to the technical field of tissue cryopreservation. Background technique [0002] Ovarian tissue freezing has its unique advantages in preserving ovarian function. First of all, the ovarian cortex contains a large number of primordial germ cells, which are located in the cortical area 1mm from the surface of the ovary. This part of the follicle is the main protection object of ovarian tissue freezing. Secondly, primordial follicles in ovarian tissue are smaller in size, in a very low metabolic state, and have very low intracellular lipid content that is sensitive to low temperature, so primordial follicles are more resistant to cryogenic freezing than mature follicles. Third, human ovarian cortex can be obtained through laparoscopy or o...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226A01N1/0284
Inventor 符晓倩
Owner 符晓倩