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Preparation and cryopreservation method and application of human placental chorionic tissues

A cryopreservation method and chorion technology, which are applied in the preservation of human or animal body, biochemical equipment and methods, and applications, can solve problems such as the influence of cell activity, loss of the function of cryopreserved tissue to protect cells, etc., to improve the activity Effect

Active Publication Date: 2018-11-16
YINFENG BIOLOGICAL GRP +1
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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
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Embodiment 1

[0037] The cryopreservation method of embodiment 1 human placenta chorionic tissue

[0038] 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.

[0039] The method of cryopreservation, the steps are as follows:

[0040] (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.

[0041] (2) The above-mentioned placental fetal chorion and large blood vessels from which the amnion has been removed are separated, washed with normal saline, and the blood vessels are flushed to remove blood clots in the blood vessels, and the...

Embodiment 2

[0048] Recovery after embodiment 2 cryopreservation

[0049] According to the method of Example 1, after cryopreservation for 6 months, recover and induce the isolation of mesenchymal stem cells, the steps are as follows: take out the cryopreserved placental chorionic tissue from liquid nitrogen, place it in the gas phase for 10 minutes to equilibrate, and quickly remove the cryopreserved Place the bag or cryopreservation tube in a water bath at 37°C to 42°C; quickly transfer the cryopreservation bag to a safety cabinet after dissolving, open the cryopreservation bag, gently remove the placental chorion with tweezers, and place it in a three-fold freezer at 4°C. In the standard concentration resuscitation solution (pre-cooled to 4°C in advance), equilibrate for 1 minute, take it out, then put it in the double standard concentration resuscitation solution at 4°C (pre-cooled to 4°C in advance), equilibrate for 1 to 3 minutes, take it out, Then put it into the resuscitation solut...

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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) separation and cryopreservation of a placental amnion and a decidua; (2) separation and cryopreservation of placenta subchorionic large vascular tissues; 3) separation and cryopreservation of the placental chorionic tissues; and (4) treatment and cryopreservation of the placental chorionic tissues. The invention also provides a method for resuscitation after cryopreservation. The cryopreservation method facilitates the improvement of the activityof cryopreserved tissues and cells, the morphology, the function and the structure of resuscitated cryopreserved tissues are same to those of fresh tissues, the overall survival rate of cells in thetissues 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 present invention relates to a preparation and cryopreservation method and application of human placental chorionic tissue, in particular to a cryopreservation method and recovery method of human placental chorionic 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...

Claims

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Application Information

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