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Systematic method for preparing and cryopreserving placental tissues according to structure layer, and application thereof

A technology of placental tissue and placenta, applied in biochemical equipment and methods, application, tissue culture, etc., can solve the problems of cell activity influence, loss of function of cryopreserved tissue protection cells, etc., and achieve the effect of promoting wound healing

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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  • Systematic method for preparing and cryopreserving placental tissues according to structure layer, and application thereof
  • Systematic method for preparing and cryopreserving placental tissues according to structure layer, and application thereof
  • Systematic method for preparing and cryopreserving placental tissues according to structure layer, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] The cryopreservation method of embodiment 1 people's placenta

[0073] 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.

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

[0075] (1) Isolation 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 side of the placenta from the placental tissue attached to it (use scissors and tweezers to carefully remove it dur...

Embodiment 2

[0101] Recovery after embodiment 2 placenta cryopreservation

[0102] 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:

[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 a safety cabinet, open the cryopreservation bag, gently remove the amniotic membrane and decidua with tweezers, put them in 4°C resuscitation solution (pre-cooled to 4°C), and equilibrate for 3 minutes; use 4°C normal saline ( (pre-cooled to 4°C in advance), washed 2 to 5 times, put in 4°C normal saline (pre-cooled to 4°C in advance), let stand, and set aside;

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

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Abstract

The invention discloses a systematic method for preparing and cryopreserving placental tissues according to a structure layer. 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 placental amnion, the decidua, the subchorionic large vascellum, the chorion and other tissues are respectively separated and preserved, so the improvement of the activity of cryopreserved tissues and cells is facilitated, 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 the tissues reaches 90% or above, and the preserved tissues can be used in the field of separation of stem cells, and also can be used in the field of tissue engineering transplantation. The systematic method for preserving various tissues of the complete placenta is provided to provide a great biological source for the studying of placenta-derived stem cells.

Description

technical field [0001] The present invention relates to a systematic method and application for preparing and freezing placental tissue according to the structure level, in particular to a method for freezing human placental amnion, chorionic blood vessels, chorion, decidua and interstitial tissues thereof, and the cryopreservation of the tissues after freezing recovery method. 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 sug...

Claims

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Application Information

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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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