Cryopreservation protection liquid for Wharton jelly tissues of human umbilical cord and preparation and application of cryopreservation protection liquid

Abstract

The invention relates to cryopreservation protection liquid for Wharton jelly tissues of a human umbilical cord. The cryopreservation protection liquid comprises 5-10 percent of a permeable cryoprotectant, 1-5 percent of a non-permeable cryoprotectant, 10-20 percent of a knockout serum replacement and 70-80 percent of a serum-free basal culture medium. The invention also discloses a preparation method of the cryopreservation protection liquid. The preparation method specifically comprises the steps of (1) precooling the serum-free basal culture medium, the non-permeable cryoprotectant and the knockout serum replacement, mixing and uniformly shaking; (2) adding the permeable cryoprotectant; and (3) performing refrigeration under a temperature condition of 2-6 DEG C for more than 30min. The invention also discloses a cryopreservation method for the Wharton jelly tissues of the human umbilical cord by the cryopreservation protection liquid. The cryopreservation method comprises the following steps of (1) pretreating the umbilical cord; (2) peeling off the Wharton jelly tissues; and (3) cryopreserving the Wharton jelly tissues of the human umbilical cord. The cryopreservation protection liquid and the cryopreservation method which are used for treating the Wharton jelly tissues of the human umbilical cord have the advantages that injury to umbilical cord mesenchymal stem cells in a cryopreservation process can be alleviated, so that the activity and the clinical use safety of the umbilical cord mesenchymal stem cells are guaranteed.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a cryopreservation solution for human umbilical cord Wharton's jelly tissue and its preparation and application. Background technique [0002] Umbilical cord mesenchymal stem cells are derived from mesoderm and ectoderm in the early stages of development. They have the potential of self-renewal, hematopoietic support, immune regulation and multi-directional differentiation. Currently, they can be used clinically for end-stage complications of diabetes, lupus erythematosus, tissue and organ damage repair And hematopoietic stem cell adjuvant therapy and many other aspects. [0003] At present, there are many kinds of stem cells in clinical application. Compared with them, umbilical cord mesenchymal stem cells have low immunogenicity, no side effects such as immune rejection in transplantation, and the advantages of simple collection process. At the same time, umbilical cord...

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Problems solved by technology

For example, the application number is 201210159916.2, and the title is "Methods for Cryopreservation, Resuscitation, Isolation and Expansion of Stem Cells of Umbilical Cord Tissue", which discloses a method for freezing and storing umbilical cord tissue. The main steps are to disinfect and clean the umbilical cord tissue, and cut the tissue into blocks; Then add the tissue block and cryopreservation solution into the cryopreservation tube, refrigerate at 4°C for 0.5h, then freeze at -80°C for 1 day, and then freeze in liquid nitrogen for later use, although It achieves the purpose of cryopreserving human umbilical cord tissue, but this clinical cryopreservation method takes more time and cost, and has the following disadvantages: 1. During the cooling process, the cryoprotective solution is at the solid-liquid phase transition point The released heat cannot be offset in time, causing the temperature of the umbilical cord tissue to rise instead, which affects the freezing effect; 2. It is impossible to accurately know the temperature change of the sample during the freezing process, which is not conducive to quality control; 3. The medium used contains animal serum components, and foreign serum components There are many uncertainties in the use, such as differences between serum batches, immunogenicity of foreign proteins, potential pathogens, etc., which may pose a threat to future clinical applications

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