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Cryopreservation method of umbilical cord tissue blocks

A technology of umbilical cord tissue and cryopreservation method, which is applied to the preservation, application, and animal husbandry of human or animal bodies. It can solve the problems of umbilical cord tissue cell death and short storage time of umbilical cord tissue, and achieve protection from damage and differentiation. Strong, simple recovery process

Active Publication Date: 2015-04-01
天晴干细胞股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims to solve the problem that the storage time of umbilical cord tissue is short and it is easy to cause the death of umbilical cord tissue cells, and provides a method for freezing umbilical cord tissue blocks

Method used

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  • Cryopreservation method of umbilical cord tissue blocks
  • Cryopreservation method of umbilical cord tissue blocks
  • Cryopreservation method of umbilical cord tissue blocks

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0019] Specific embodiment one: the present embodiment umbilical cord tissue block cryopreservation method, carry out according to the following steps:

[0020] 1. Processing of organization blocks:

[0021] In the aseptic workbench, the umbilical cord was disinfected with alcohol with a concentration of 75% by volume, and then washed twice with normal saline. Peel off the Huatong glue, put the Huatong glue in a clean centrifuge tube, and cut it to 1-2mm with surgical scissors 3 blocky;

[0022] 2. Freezing of tissue blocks:

[0023] Add DMEM to the tissue block. The volume of DMEM is half of the volume of the tissue block. After resuspending the tissue block, add pre-frozen storage solution equal to the volume of DMEM, and then distribute 1 mL per tube into cryopreservation tubes. The program-controlled cooling device lowers the temperature, specifically: first balance at 4°C for 30 minutes, then drop to -5°C at a rate of 1°C / min, then drop to -54°C at a rate of 21°C / min, ...

specific Embodiment approach 2

[0025] Embodiment 2: This embodiment is different from Embodiment 1 in that: the cleaned umbilical cord is cut into sections with a length of 2.5 cm. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0026] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is: use surgical scissors to cut to 1.5mm 3 blocky. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a cryopreservation method of umbilical cord tissue blocks, and relates to a cryopreservation method of umbilical cords. The invention aims at solving the problems that the preservation time of umbilical cord tissues is short, and the cell death of the umbilical cord tissues is easily caused at present. The method comprises the following steps: 1, carrying out treatment of the tissue blocks, namely in a sterile operating platform, carrying out disinfection treatment on an umbilical cord, cleaning the umbilical cord normal saline, shearing the umbilical cord into segments, peeling off a wharton jelly, placing the wharton jelly in a clean centrifuging tube and cutting the wharton jelly into pieces; and 2, adding DMEM in the tissue blocks, resuspending the tissue blocks, then adding a pre-cooling cryopreservation liquid, sub-packaging the tissue blocks in cryopreservation tubes, cooling by use of a program-controlled cooler, and transferring the tissue blocks in liquid nitrogen for long-time storage after bacterial detection, microbiological detection and virus detection are all negative, so as to finish the cryopreservation of the umbilical cord tissue blocks. The directly-inoculated adherent survival rate of the cryopreserved and unfrozen tissue blocks in culture bottles is more than 98%, the cell growth is good, and moreover, an osteogenic differentiation experiment carried out on the cultured cells of the cryopreserved tissue blocks indicates that the differentiation function is high. The method is used for storing the umbilical cord tissues.

Description

technical field [0001] The invention relates to a freezing method for umbilical cord. Background technique [0002] The umbilical cord is a tubular structure connecting the fetus and the placenta. It has two arteries and one vein. There is a loose jelly-like substance (called Wharton's jelly) between the blood vessels and the inner wall. The jelly-like substance is rich in umbilical cord mesenchyme. stem cells. [0003] Mesenchymal stem cells are important members of the stem cell family. They are primitive cells with self-replication and multi-directional differentiation potential. They are the origin cells of the body and the primitive cells that form various tissues and organs of the human body. Under certain conditions, it can differentiate into a variety of functional cells or tissues and organs, which are called "universal cells" in the medical field, and are suitable as seed cells for the repair of tissue and organ damage caused by aging and disease. [0004] Stem c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 王灵娟张怡王浩宇梁丽潮王博昊赵新新
Owner 天晴干细胞股份有限公司
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