BEIJING OBSTETRICS & GYNECOLOGY HOSPITAL CAPITAL MEDICAL UNIV
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In developed countries in Europe and the United States, ovarian tissue cryopreservation and retransplantation have been used as routine fertility protection techniques for more than ten years, but there is no live birth report in my country
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Embodiment 1-6
[0099] Embodiment 1-6 Preparation of Ovarian Tissue Cryopreservation Solution
[0100] Take the preparation of 100ml cryopreservation solution as an example: Measure LeibovitzL-15 medium, DMSO, HSA, and SSS into the reagent bottle according to the amount of each component in Table 1, shake gently to mix, and use a 0.22um filter to Sterilize by filtration, filter into an autoclaved reagent bottle, store at 4°C for later use, or freeze at -20°C, and move to a 4°C refrigerator for later use.
[0101] Table 1 Components of Cryopreservation Solution 1
[0102]
Embodiment 7-17
[0103] Example 7-17 Preparation of Ovarian Tissue Cryopreservation Solution
[0104] Take the preparation of 100ml cryopreservation solution as an example: Measure LeibovitzL-15 medium, DMSO, HSA, and SSS into the reagent bottle according to the amount of each component in Table 2, shake gently to mix, and then add the corresponding amount taurine, nitrogen acetylcysteine, vitamin E, vitamin C and heparin. After the solid is completely dissolved, use a 0.22um filter to filter and sterilize, filter into an autoclaved reagent bottle, store at 4°C for later use, or freeze at -20°C, and move to a 4°C refrigerator for later use.
[0105] Table 2 Components of cryoprotective solution 2
[0106]
[0107]
Embodiment 18-20
[0108] The preparation of embodiment 18-20 ovarian tissue resuscitating liquid
[0109] Take the preparation of 100ml cryopreservation solution as an example: Measure LeibovitzL-15 medium, DMSO, HSA, and SSS into the reagent bottle according to the amount of each component in Table 2, shake gently to mix, or add the corresponding Amount of taurine, nitrogen acetylcysteine, vitamin E, vitamin C. After the solid is completely dissolved, use a 0.22um filter to filter and sterilize, filter into an autoclaved reagent bottle, store at 4°C for later use, or freeze at -20°C, and move to a 4°C refrigerator for later use.
[0110] Table 3 Composition of resuscitation solution
[0111]
[0112]
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Abstract
The invention discloses a cryopreservation resuscitation fluid for ovarian tissues. The resuscitation fluid comprises the following components: (1) a resuscitation solution 1: 90V / V% DPBS, 10V / V% HSA, 0.5-0.8 mol / L sucrose and 5mM N-acetylcysteine; (2) a resuscitation solution 2: 90 V / V% DPBS, 10 V / V% HSA, 0.3-0.5 mol / L sucrose, and 5 mM N-acetylcysteine; and (3) a resuscitation solution 3: 90 V / V% DPBS, 10 V / V% HSA, 0.12-0.3 mol / L sucrose, and 5 mM N-acetylcysteine; during resuscitation, the method comprises the following steps: (1) taking out the cryopreservation tube filled with the ovarian tissue from the liquid nitrogen, and dissolving; and (2) taking out the ovarian tissue from the cryopreservation tube, sequentially adding the ovarian tissue into solutions of the resuscitation solution 1, the resuscitation solution 2 and the resuscitation solution 3, respectively keeping for a certain time, and then putting the ovarian tissue into an eluent for elution to finish resuscitation. The cryopreservation resuscitation fluid can improve the survival rate of resuscitated ovarian tissues.
Description
[0001] This application is a divisional application of Chinese patent application 201810254362.1. [0002] Original application date: 2018.3.26 [0003] Original application number: 201810254362.1 [0004] Invention name of the original application: a cryopreservation protection and resuscitation solution for ovarian tissue technical field [0005] The invention belongs to the field of cryopreservation of ovarian tissue, in particular to a cryopreservation recovery solution of ovarian tissue. Background technique [0006] 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 happin...
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