Preparation and cryopreservation method and application of human placental villous tissue

A cryopreservation method and technology of human placenta, which is applied in the field of preparation and cryopreservation of human placental villi tissue, can solve the problems of losing the function of protecting cells of cryopreserved tissue and the influence of cell activity

Active Publication Date: 2021-07-23
YINFENG BIOLOGICAL GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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.

Method used

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  • Preparation and cryopreservation method and application of human placental villous tissue
  • Preparation and cryopreservation method and application of human placental villous tissue
  • Preparation and cryopreservation method and application of human placental villous tissue

Examples

Experimental program
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Effect test

Embodiment 1

[0039] The cryopreservation method of embodiment 1 people's placenta villous tissue

[0040] 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.

[0041] The method of cryopreservation, the steps are as follows:

[0042] (1) Wash the placenta (to remove dirt and microbial contamination); cut off the decidua along the edge of the placenta to remove the amniotic membrane.

[0043] (2) Separate the above-mentioned placental chorion and large blood vessels from which the amniotic membrane has been removed, and the remaining placental tissue is the placental villi tissue, and cut the placental villi tissue into 5 cm ...

Embodiment 2

[0052] Recovery after embodiment 2 cryopreservation

[0053] 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:

[0054] (2) Recovery of cryopreserved placental villus tissue: Take out the cryopreserved placenta villi tissue from liquid nitrogen, place the cryopreserved bag in the gas phase for 10 minutes to equilibrate, and quickly place the cryopreservation bag in a water bath at 37°C to 42°C; Transfer the cryopreservation bag to a safety cabinet, open the cryopreservation bag, gently remove the placental villus tissue with tweezers, put it in a resuscitation solution with three times the reference concentration at 4°C (pre-cooled to 4°C in advance), equilibrate for 1 minute, and take it out. Then put it into the resuscitation solution of double the standard concentration at 4°C (pre-cooled to 4°C in advance), equilibrate for 1 to 3 minutes, take...

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Abstract

The invention discloses a method for preparing and freezing human placental villi, comprising: (1) washing the placenta; cutting off the decidua along the edge of the placenta to remove the amnion; (2) removing the placenta from which the amnion has been removed The fetal facial chorion and large blood vessels are separated together, and the remaining placental tissue is the placental villi tissue. Cut the placental villi tissue into small pieces, and then cut them into thin slices; put the thin slices into cryopreservation bags or tubes, and proceed in three steps. Introduce the vitrification solution by using the method, transfer to the program cooling device, cool down to -80°C to -90°C, 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 those of fresh tissues, and the overall survival rate of cells in the tissues is over 90%, and the preserved tissues can not only be used to separate stem cells , epithelial cells and other fields, and 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 villus tissue and its application, in particular to a method for cryopreserving and resuscitating the human placenta villi 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 membrane-derived mesenchymal stem cells, but also to be used in ophthalm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02C12N5/073
CPCA01N1/0221C12N5/0605C12N2500/32C12N2500/34
Inventor 崔光晶徐峰波金华宋现收生德伟李德柱
Owner YINFENG BIOLOGICAL GRP
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