Preparation and cryopreservation method and application of human placental amniotic membrane and decidua tissue
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A technology of decidual tissue and cryopreservation method, which is applied in the preservation of human or animal body, biochemical equipment and methods, and application, etc. active effect
Active Publication Date: 2021-11-26
YINFENG BIOLOGICAL GRP
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Therefore, if the volume of cryopreserved tissue is too small, the cells will be more exposed to the toxic components of the cryoprotectant, the cell viability will be affected, and the function of cryopreserved tissue to protect cells will be lost.
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Embodiment 1
[0029] The cryopreservation method of embodiment 1 human placenta amniotic membrane, decidua
[0030] Placenta collection: Select a healthy placenta without infectious diseases and obstetric complications, obtain the consent of the parturient and sign the informed consent; for normal collection, the collected placenta will be transported to the laboratory within 48 hours, and various necessary tests will be carried out, such as Detection of infectious diseases such as viruses, detection of bacterial contamination, etc.
[0031] The method of cryopreservation, the steps are as follows:
[0032] (1) Wash the placenta (to remove dirt and microbial contamination); cut off the decidua along the edge of the placenta, and divide the amniotic membrane into 10cm 2 ~100cm 2 Then, remove the amniotic membrane on the fetal side of the placenta from the placental tissue attached to it (use scissors and tweezers to carefully remove it during the specific operation, pay attention to keep t...
Embodiment 2
[0036] Recovery after embodiment 2 cryopreservation
[0037] According to the method of Example 1, after cryopreservation for 6 months, recovery was carried out, and mesenchymal stem cells were induced and isolated. The steps were as follows:
[0038] Take out the cryopreserved placental amniotic membrane and decidua from the liquid nitrogen, and quickly place the cryopreservation bag in a water bath at 37°C to 42°C; quickly transfer the cryopreservation bag to a safety cabinet after dissolution, open the cryopreservation bag, Gently remove the amniotic membrane and decidua with tweezers, put them in 4°C resuscitation fluid (pre-cooled to 4°C in advance), and equilibrate for 3 minutes; rinse with 4°C normal saline (pre-cooled to 4°C in advance) for 2 to 5 times, put Put it into 4°C physiological saline (pre-cooled to 4°C), let it stand, and set aside;
[0039] The resuscitation solution is composed of trehalose and hydroxyethyl starch sodium chloride injection, wherein trehal...
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Abstract
The invention discloses a method for preparing and freezing human placental amniotic membrane and decidua tissue, comprising: (1) washing the placenta; cutting off the decidual membrane along the edge of the placenta, and dividing the amniotic membrane into tissue pieces; The amniotic membrane on the fetal surface of the placenta is removed from the attached placental tissue; (2) Wash the amniotic membrane and decidua with normal saline or PBS buffer solution, fold or lay the amniotic membrane and decidua into a cryopreservation bag or cryopreservation tube , add 4°C cryopreservation solution, seal, transfer to a programmed cooling device, cool down to -80°C according to the set cooling program, and transfer to liquid nitrogen for cryopreservation. The cryopreservation method of the present invention is beneficial to improve the activity of cryopreserved tissues and cells, and the shape, function and structure after recovery are consistent with fresh tissues, and the overall survival rate of cells in the tissues is over 90%, and the preserved tissues can not only be used for separation Stem cells, epithelial cells and other fields can also be used in fields such as tissue engineering and transplantation.
Description
technical field [0001] The invention relates to a method for preparing and freezing human placental amniotic membrane and decidua tissue and an application thereof, in particular to a freezing preservation method and a resuscitation method for human placental amniotic membrane and decidua tissue. Background technique [0002] The placenta is an important organ for material exchange between the fetus and the mother. It is a tissue-combining organ between the mother and the child formed by the joint growth of the embryonic embryonic membrane and the maternal endometrium during human pregnancy. In recent years, with the deepening of scientific research, people can classify various types of cells from placenta, including hematopoietic stem cells, mesenchymal stem cells, epithelial cells, etc., thus changing the long-term status of placenta as medical waste. [0003] Modern scientific research suggests that placental amniotic membrane can be used not only to expand amniotic membr...
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