Cell freezing medium, freezing method for mesenchymal stem cells and application of cell freezing medium

Abstract

The invention provides a cell freezing medium which comprises the following components in percentage by volume: 1%-10% of a dimethyl sulfoxide solution, 1%-10% of a hydroxyethyl starch sodium chloride solution, 5%-20% of a human serum albumin solution, 2%-20% of a glucose solution, 5%-20% of a dextran 40 sodium chloride solution, 0.1%-1% of a non-essential amino acid solution and the balance of Hank's a balanced salt solution, and the sum of the above components is 100%. The cell freezing medium is simple in composition and convenient to prepare, mostly adopts medicinal-grade auxiliary materials, can meet the requirements of declaration of cell treatment drugs, is good in freezing effect, can stably keep the recovery rate of frozen cells to be 90% or above, can stably keep the recovery rate of the recovered cells to be 90% or above, is good in freezing effect, and provides a guarantee for patent medicines for cell treatment.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a cell cryopreservation solution and a cryopreservation method for mesenchymal stem cells. Background technique [0002] Stem cells have been widely used in scientific research and clinical treatment due to their self-renewal and multi-directional differentiation potential. Mesenchymal stem cells in stem cells have been widely used in therapeutic research of animal disease models and human clinical experiments. Mesenchymal stem cells Diseases that can be treated include diabetes, liver cirrhosis, skin damage, myocardial infarction, dry eye disease, and nerve damage. In addition, mesenchymal stem cells also have the functions of promoting hematopoiesis and immune regulation, and are currently used worldwide. There are stem cell drug products on the market. [0003] As an effective method to preserve cells, cell cryopreservation technology has been widely used in the field of ...

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Problems solved by technology

[0003] As an effective method to preserve cells, cell cryopreservation technology has been widely used in the field of biology, because under normal circumstances, direct freezing of cells will produce ice crystals that will cause damage to cells, resulting in cell death, so cryopreservation of cells It is necessary to add a certain proportion of cryopreservation solution to maintain the integrity of the cells after freezing and thawing and minimize the damage to the cells, but it is inevitable that the activity of the cells will be reduced and the cells will be damaged during the cryopreservation process.

[0004] At present, there are many patent technologies on cell cryopreservation solutions, but each has its own technical characteristics. The most commonly used cryopreservation solution is usually fetal bovine serum (FBS) with 10% dimethyl sulfoxide (DMSO) solution, DMSO can Reduce the formation of ice crystals, reduce the damage of free radicals to cells, and change the permeability of biofilms to electrolytes, drugs, poisons and metabolites, which can protect cells well during cell cryopreservation, but the use of FBS has the potential for animal-derived viruses and Cytokines and other potential safety hazards are not conducive to the scale and industrialization of cell products, thus limiting the further improvement of this technology

Human platelet lysate (HPL) is also used instead of traditional FBS to become animal-free cell cryopreservation solution, but the long-term cryopreservation effect of this type of cell cryopreservation solution is not good. At the same time, different batches of human platelet lysate (HPL ) are different, resulting in unstable quality of frozen cells

Some patented technologies are based on the fact that they can be directly used for human infusion. The ingredients used in this type of cryopreservation solution are relatively simple. Generally, a small amount of DMSO and albumin are added to buffered saline solution. The formula of this type of cryopreservation solution is simple. But the cryopreservation effect is not ideal, the survival rate is difficult to maintain above 90% stably, the recovery rate and the adherence rate are even worse, and the cryopreservation process is relatively demanding, and a specific cryopreservation program needs to be developed for it; some patents In order to improve the effect of cryopreservation, some small molecular substances are added, such as insulin, heparin, VEGF, HGF, etc. Although the addition of these small molecules may improve the effect of cell cryopreservation, the addition of these small molecular substances increases the use of cells. Risks to humans, especially hormones

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