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Preparation and cryopreservation method and application of human placental chorionic tissues

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: 2018-11-16
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 chorionic tissues
  • Preparation and cryopreservation method and application of human placental chorionic tissues
  • Preparation and cryopreservation method and application of human placental chorionic tissues

Examples

Experimental program
Comparison scheme
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 preparation and cryopreservation method of human placental chorionic tissues. The method comprises the following steps: (1) cleaning a placenta, and cutting off a shed decidua along the edge of the placenta to remove the amnion; and (2) separating the amnion-removed placenta fetus chorion and large vessels to obtain remaining placenta tissues which are placental chorionictissues, cutting the placental chorionic tissues to form small blocks, cutting the small blocks to form slices, placing the slices in a cryopreservation bag or a cryopreservation tube, guiding a vitrification cryopreservation solution through a three-step process, transferring the obtained mixture into a programmable cooler, reducing the pressure to -80 to -90 DEG C, transferring the cooled mixture to liquid nitrogen, and performing cryopreservation. The cryopreservation method facilitates the improvement of the activity of cryopreserved tissues and cells, the morphology, the function and thestructure of resuscitated cryopreserved tissues are same to those of fresh tissues, the overall survival rate of cells in the tissues reaches 90% or above, and the preserved tissues can be used in the field of separation of stem cells and epithelial cells, and also can be used in the field of tissue engineering 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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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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