Cryopreservation and resuscitation method for ovarian tissue
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A technology of ovarian tissue and cryopreservation method, which is applied in the direction of tissue culture, preservation of human or animal body, biochemical equipment and methods, etc., to achieve the effect of good cryopreservation of fertility, ensuring uniformity, and good cell membrane structure
Active Publication Date: 2018-10-12
阮祥燕
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In developed countries in Europe and the United States, ovarian tissue cryopreservation and retransplantation have been
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[0096] Example 1-6 Preparation of ovarian tissue cryopreservation protection solution
[0097] Take the preparation of 100ml cryopreservation solution as an example: Measure LeibovitzL-15 medium, DMSO, HSA, SSS and add them to the reagent bottle according to the amount of each component in Table 1. Shake gently to mix, use a 0.22um filter Sterilize by filtration. Store the filter paper in the autoclaved reagent bottle at 4°C for later use, or freeze at -20°C, and move to 4°C refrigerator for later use.
[0098] Table 1 Composition of cryopreservation solution 1
[0099]
Example Embodiment
[0100] Example 7-17 Preparation of ovarian tissue cryopreservation protection solution
[0101] Take the preparation of 100ml cryopreservation solution as an example: Measure LeibovitzL-15 medium, DMSO, HSA, SSS according to the amount of each component in Table 2 and add them to the reagent bottle, shake gently to mix, and then add the corresponding amount Of taurine, nitrogen acetylcysteine, vitamin E, vitamin C. After the solid is completely dissolved, filter and sterilize with a 0.22um filter. The filter paper is autoclaved in a reagent bottle and stored at 4°C for later use, or frozen at -20°C, and moved to a 4°C refrigerator for later use.
[0102] Table 2 Composition of cryopreservation solution 2
[0103]
[0104]
Example Embodiment
[0105] Example 18-20 Preparation of ovarian tissue resuscitation solution
[0106] Take the preparation of 100ml cryopreservation solution as an example: according to the amount of each component in Table 2, measure LeibovitzL-15 medium, DMSO, HSA, SSS into the reagent bottle, shake gently to mix, or add the corresponding The amount of taurine, nitrogen acetylcysteine, vitamin E, vitamin C. After the solid is completely dissolved, filter and sterilize with a 0.22um filter. The filter paper is autoclaved in a reagent bottle and stored at 4°C for later use, or frozen at -20°C, and moved to a 4°C refrigerator for later use.
[0107] Table 3 Composition of resuscitation fluid
[0108]
[0109]
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Abstract
The invention discloses a cryopreservation and resuscitation method for an ovarian tissue. The cryopreservation and resuscitation method for the ovarian tissue comprises a cryopreservation method anda resuscitation method. The cryopreservation method comprises the following steps that (1) pretreatment is conducted, specifically, the ovarian tissue is placed in a transfer liquid, and is cut into tissue slices with medulla removed and cortex reserved; (2) osmotic balance is conducted, specifically, the tissue slices are transferred into a sample box containing a cryopreservation protective liquid and then placed in ice for balancing; and (3) cryopreservation is conducted, specifically, a certain amount of cryopreservation protective liquid is added into a cryopreservation tube, a tissue block is placed in the cryopreservation tube, then gradual temperature decreasing is conducted, and liquid nitrogen cryopreservation is conducted. According to the cryopreservation and resuscitation method, damage to the ovarian tissue can be reduced through the liquid nitrogen cryopreservation method with gradual temperature decreasing, meanwhile, meanwhile the cryopreservation protective liquid anda resuscitation protective agent are used cooperatively, the ovarian tissue can be cryopreserved quickly and efficiently, and the viability of the ovarian tissue after resuscitation can be increased.
Description
technical field [0001] The invention belongs to the field of cryopreservation of ovarian tissue, and specifically relates to a method for cryopreservation and recovery of ovarian tissue, and simultaneously relates to a cryopreservation protection solution and a recovery solution. Background technique [0002] There are about 4.3 million new cancer patients in China every year, and the five-year survival rate of cancer patients can be as high as 80-90%, and more than 70% of the survivors have urgent needs for fertility. However, while saving the lives of cancer patients, radiotherapy and chemotherapy have caused 70%-100% of the ovarian function is damaged, making it lose fertility, which greatly affects the patient's physical and mental health and family happiness; due to the early decline of ovarian function, various chronic diseases such as cardiovascular disease, diabetes, bone disease, etc. Poor disease, Alzheimer's disease and early death rate are significantly increased...
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