Umbilical cord tissue cryopreservation liquid and preparation method thereof, and cryopreservation method and culture method of umbilical cord tissue

Abstract

The invention relates to an umbilical cord tissue cryopreservation solution and a preparation method thereof, and a cryopreservation method and a culture method of umbilical cord tissues. The umbilical cord tissue cryopreservation liquid is prepared from human autoserum, human serum albumin and auxiliaries, wherein the auxiliary agent is composed of at least three of vitamin K2, trehalose, EGF and BFGF. The preparation method comprises the following steps: uniformly mixing the human autoserum, the 20% human serum albumin and the auxiliary agent to obtain the umbilical cord tissue freezing medium; then freezing the umbilical cord tissue by using an umbilical cord tissue freezing medium; and taking out the cryopreserved umbilical cord tissue from the liquid nitrogen, thawing the tissue in a water bath at 37 DEG C, and performing cell culture. The invention aims to solve technical problem of how to provide the umbilical cord tissue cryopreservation liquid, so that the umbilical cord tissue cryopreserved by the cryopreservation liquid can be directly subjected to cell culture after being thawed without being cleaned. The umbilical cord tissue cryopreserved by the cryopreservation medium is high in multiplication capacity, the number of primary cells cultured by the cryopreservation medium is large, the motility rate is high, and the multiplication capacity cannot be attenuated along with the increase of the cryopreservation time. The method is economical in cost, fast, efficient and more practical.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an umbilical cord tissue cryopreservation liquid and a preparation method thereof, a cryopreservation method and a culture method of umbilical cord tissue. Background technique [0002] The Wharton's jelly in the umbilical cord is rich in umbilical cord mesenchymal stem cells. 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. Stem cells are a type of undifferentiated and immature ce...

AI Technical Summary

Problems solved by technology

[0005] Umbilical cord tissue cryopreservation solutions in the prior art have the following defects in cryopreservation: one is that some of the existing cryopreservation solutions contain DMSO, some contain exogenous / animal-derived serum components, or add some potentially risky Therefore, the cryopreservation solution has a potential safety risk to the frozen umbilical cord tissue; the other is that the components of the cryopreservation solution have potential safety hazards, so the umbilical cord tissue needs to be washed repeatedly before tissue culture , the operation is cumbersome and the efficiency is low; the third is that the umbilical cord tissue frozen in the existing cryopreservation solution has poor proliferation ability in tissue culture due to the influence of the components of the cryopreservation solution on the umbilical cord tissue; the fourth is that the existing cryopreservation solution The main raw materials of the liquid need to be purchased, and the umbilical cord tissue needs to be repeatedly cleaned after thawing and thawing, and the cost is high

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