Preservation solution for umbilical mesenchymal stem cells

Abstract

The invention provides a preservation solution for umbilical mesenchymal stem cells and a preparation method for the preservation solution and belongs to the technical field of stem cell preservation.The cell preservation solution provided by the invention comprises 85%-93% (v / v) of fetal calf serum, 5%-12% (v / v) of dimethyl sulfoxide, 0.4%-0.8% (v / v) of low-molecular-weight calcium heparin injection, 1%-1.2% (v / v) of low-molecular-weight sodium heparin injection, 0.2%-0.4% (w / v) of anthocyan and 0.4%-0.6% (w / v) of epicatechin. The umbilical mesenchymal stem cells are subjected to separation,culture and subculture and then are preserved in the preservation solution, so that the activity and survival rate of the stem cells can be maintained, meanwhile, the stability of preservation can beimproved, the preservation time can be prolonged, and the risk can be lowered.

Description

technical field [0001] The invention relates to the technical field of stem cell preservation, in particular to a storage solution of umbilical cord mesenchymal stem cells. Background technique [0002] Umbilical cord blood is the blood remaining in the placenta and umbilical cord after the fetus is delivered, the umbilical cord is ligated and severed. Umbilical cord mesenchymal stem cells refer to a kind of multifunctional stem cells present in the umbilical cord tissue of newborns, which can differentiate into many kinds of tissue cells. It has broad clinical application prospects. Usually, umbilical cord mesenchymal stem cells will continue to divide, grow, and then age when cultured in vitro, resulting in loss of function and loss of multiple differentiation potentials. Therefore, when the prepared stem cells cannot be used immediately, they need to be stored. Liquid has a great impact on the activity and survival rate of cells, which will further affect the safety and ...

AI Technical Summary

Problems solved by technology

Usually, umbilical cord mesenchymal stem cells will continue to divide, grow, and then age when cultured in vitro, resulting in loss of function and loss of multiple differentiation potentials. Therefore, when the prepared stem cells cannot be used immediately, they need to be stored. Liquid has a great impact on the activity and survival rate of cells, which will further affect the safety and effectiveness of clinical treatment

[0003] At present, normal saline is mostly used in the storage solution of umbilical cord mesenchymal stem cells, but the normal saline is acidic. At the same time, the activity and survival rate of the cells are poor when this kind of storage solution is used to store the cells. It is only suitable for short-term storage of cells. suitable for long-term storage

In addition, dimethyl sulfoxide and fetal bovine serum are also very commonly used cell storage solutions. Among them, dimethyl sulfoxide can prevent cell frostbite, but it has certain toxicity to cells, causing damage to stem cells and affecting the storage effect. Fetal bovine serum is often added as a protective agent to reduce the toxicity of dimethyl sulfoxide to cells and improve cell survival, but animal serum often has the risk of bacterial infection, leading to adverse reactions. In addition, fetal bovine serum is different due to different batches. There is also the problem of instability

[0004] Chinese patent CN201510518785.6 discloses a serum-free preservation solution for umbilical cord mesenchymal stem cells and its application. The preservation solution in this invention includes sodium chloride and normal saline, and insulin, estrogen and progesterone added to the normal saline, Hydrocortisone, sodium selenite, adenosine, N-acetylcysteine ​​and L-glutamine, this preservation solution can provide active nutrients and energy substances, maintain a low metabolic level, and at the same time, avoid growth However, when the preservation solution is used at a lower temperature, although there are nutrients and energy substances provided, the cells cannot be preserved due to frostbite and activity damage. A certain level of metabolism, which in turn leads to lower cell viability

However, this paper only compared different preservation solutions to investigate the effects of preservation solutions and storage time on the survival rate of umbilical cord-derived mesenchymal stem cells, and did not further improve and optimize the preservation solutions to address the problems existing in the existing technology

[0006] In view of the low cell survival rate, low cell activity, high risk of adverse reactions and poor stability in the storage solution of umbilical cord mesenchymal stem cells, it is necessary to find a storage solution of umbilical cord mesenchymal stem cells so that the umbilical cords preserved in the storage solution Mesenchymal stem cells have high cell survival rate and high activity after resuscitation, while reducing the occurrence of adverse reactions, improving stability and prolonging storage time

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