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A systematic method and application of preparation and cryopreservation of placental tissue according to the structure level

A placental tissue, cryopreservation technology, applied in the recovery after tissue cryopreservation, in the field of cryopreservation of human placental amniotic membrane, chorionic blood vessels, chorion and decidua and its interstitial tissue, can solve the effect of cell activity, loss of cryopreservation The role of tissue protection cells and other issues to achieve the effect of improving 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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  • A systematic method and application of preparation and cryopreservation of placental tissue according to the structure level
  • A systematic method and application of preparation and cryopreservation of placental tissue according to the structure level
  • A systematic method and application of preparation and cryopreservation of placental tissue according to the structure level

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Embodiment 1 The cryopreservation method of human placenta

[0073] Placenta collection: select healthy placenta without infectious diseases and obstetric complications, and obtain the consent of the mother and sign the informed consent; normal collection, transport the collected placenta to the laboratory within 48 hours, and carry out various necessary tests, such as Virus and other infectious diseases detection, bacterial contamination detection, etc.

[0074] The method of cryopreservation, the steps are as follows:

[0075] (1) Separation and cryopreservation of placental amniotic membrane and decidua: 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 surface of the placenta from the attached placental tissue (use scissors and tweezers to carefully remove it during the specific operation, pay atten...

Embodiment 2

[0101] Example 2 Recovery of placenta after cryopreservation

[0102] After 6 months of cryopreservation according to the method of Example 1, recovery was performed, and the mesenchymal stem cells were induced and isolated, and the steps were as follows:

[0103] (1) Recovery of cryopreserved placental amniotic membrane and decidua: Take out the cryopreserved placental amniotic membrane and decidua from liquid nitrogen, and quickly place the cryopreservation bag in a water bath at 37°C to 42°C; Transfer the storage bag to the safety cabinet, open the freezing bag, gently remove the amniotic membrane and decidua with tweezers, put them in the resuscitation solution at 4 °C (pre-cooled to 4 °C in advance), and equilibrate for 3 minutes; (pre-cooled to 4°C in advance), rinsed 2 to 5 times, put into normal saline at 4°C (pre-cooled to 4°C in advance), and set aside for later use;

[0104] The resuscitation liquid is composed of trehalose and hydroxyethyl starch sodium chloride i...

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Abstract

The invention discloses a systematic method for preparing and freezing placental tissue according to structural levels, including: (1) separation and freezing of placental amniotic membrane and decidua; (2) separation and freezing of placental subchorionic large blood vessel tissue ; (3) Isolation and cryopreservation of placental chorionic tissue; (4) Processing and cryopreservation of placental chorionic tissue. The invention also provides a recovery method after cryopreservation. The present invention separates and preserves tissues such as placental amniotic membrane, decidua, subchorionic blood vessels, and chorion, which is beneficial to improving the activity of frozen tissues and cells. The overall survival rate of inner cells is over 90%, and the preserved tissues can not only be used in fields such as separating stem cells, but also in fields such as tissue engineering and transplantation. The invention provides a systematic method for preserving various tissues of the complete placenta, and provides important biological resources for the research on stem cells derived from the placenta.

Description

technical field [0001] The invention relates to a system method and application for preparing placental tissue according to structural levels and cryopreserving it, in particular to a cryopreservation method for human placental amniotic membrane, chorionic blood vessels, chorion and decidua and their interstitial tissues, and the cryopreserved tissue after cryopreservation. recovery method. Background technique [0002] The placenta is an important organ for material exchange between the fetus and the mother, and is a tissue-binding organ between the mother and the child formed by the joint growth of the 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 the placenta, including hematopoietic stem cells, mesenchymal stem cells, and epithelial cells, thus changing the long-term status of the placenta as a medical waste. [0003] Modern scientific re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02C12N5/073C40B50/06
CPCA01N1/0221C12N5/0605C12N2500/32C12N2500/34C40B50/06
Inventor 徐峰波王肇光崔光晶金华宋现收生德伟李德柱
Owner YINFENG BIOLOGICAL GRP
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