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A technology of ovarian tissue and preservation method, applied in the field of reproductive medicine, can solve problems such as cytotoxic damage
Inactive Publication Date: 2014-12-24
THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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[0006] The purpose of the present invention is to provide a method for deep cryopreservation of ovarian tissue to solve th...
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Embodiment 1
[0033] For the cryopreservation method of ovarian tissue provided by the embodiment of the present invention, please refer to figure 1 , including the following steps:
[0034] Step 101: Incubate the ovarian tissue to be frozen with a hyaluronidase solution with a volume concentration of 8-12% at room temperature for 10-15 minutes to obtain the ovarian tissue after enzymatic hydrolysis;
[0035] In step 101, which is the most critical step of the present invention, after the ovarian tissue to be frozen is incubated at room temperature with 8-12% hyaluronidase solution for 10-15 minutes, the hyaluronidase can detoxify the collagen contained in the ovarian tissue, etc. The tissue is subjected to enzymatic hydrolysis. After enzymatic hydrolysis, the overall structure of the ovarian tissue is loose. Therefore, it is very conducive to the penetration of the cryoprotectant in the subsequent pre-equilibration process, and directly shortens the time of the pre-equilibration process. ...
Embodiment 2
[0043] Please refer to figure 2 , in this embodiment, the preparation method of ovarian tissue cryopreservation method comprises the following steps:
[0044] Step 201: collecting ovarian tissue and placing it in L-15 culture solution containing 10% fetal bovine serum;
[0045] In this step, the ovarian tissue is placed in L-15 culture solution with 10% fetal bovine serum to ensure its nutrient supply, thereby ensuring its tissue activity. In addition, the L-15 culture solution with 10% fetal bovine serum realizes The effect of blood supply provides sufficient nutrition for the normal culture of ovarian tissue.
[0046] Step 202: Transfer the L-15 culture solution with ovarian tissue to an ice box, transfer it to a sterile ultra-clean workbench within 8-12 minutes, and replace it with fresh L-15 culture solution containing 10% fetal bovine serum ;
[0047] In step 202, in order to avoid tissue ischemia damage caused by contamination and long in vitro time, the time for tra...
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Abstract
The invention relates to the field of reproductive medicines, in particular to an ovarian tissue cryopreservation method. The ovarian tissue cryopreservation method comprises the following steps: culturing an ovarian tissue to be frozen by a hyaluronidase solution with the volume concentration of 8-12 percent for 10-15 minutes so as to obtain an enzymolyzed ovarian tissue; adding the enzymolyzed ovarian tissue into a freezing protection agent for prebalancing under the temperature of 3-5 DEG C for 10-15 minutes to obtain a prebalanced ovarian tissue; performing programmed cryopreservation on the prebalanced ovarian tissue to reduce the temperature of the prebalanced ovarian tissue to be minus 150 DEG C, and transferring the prebalanced ovarian tissue into a liquid nitrogen tank for storage. Compared with the common prebalancing time of 30-90 minutes in the prior art, the prebalancing time of the ovarian tissue cryopreservation method is greatly shortened, so that the cytotoxic damage, which is caused by the freezing protection agent, to the ovarian tissue under overlong prebalancing time is avoided.
Description
technical field [0001] The invention relates to the field of reproductive medicine, in particular to a method for cryopreservation of ovarian tissue. Background technique [0002] In recent years, the incidence of malignant tumors has shown a trend of increasing incidence and younger age. With the advancement of early diagnosis and radiotherapy and chemotherapy, the cure rate of various malignant tumors can reach more than 90%. However, after childhood, adolescence, and women of childbearing age receive high-dose combined chemotherapy and radiotherapy, the gonadal toxicity may lead to premature ovarian failure and loss of fertility, which greatly reduces the quality of life of female patients who survive long-term after treatment, and seriously affects their society. family role. Therefore, the protection and preservation of female fertility has become an urgent issue in the field of reproductive medicine. In the preservation of female fertility, cryopreservation of ovaria...
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IPC IPC(8): A01N1/02
Inventor 肖准
Owner THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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