Umbilical cord blood hematopoietic stem cell preservation solution and preparation method thereof

Abstract

The invention provides a cord blood hematopoietic stem cell preservation solution and a preparation method thereof, belonging to the technical field of cell preservation. The raw materials of the preservation solution include: low molecular weight dextran, dimethyl sulfoxide, DMEM medium, glycerol, folic acid, mannose Polysaccharide and phosphatidylserine are mixed with raw materials to obtain a preservation solution, and then mixed with hematopoietic stem cells after two stages of variable temperature pretreatment to preserve hematopoietic stem cells. The prepared preservation solution has a long preservation time, the preserved stem cells have less cell damage after resuscitation, and the survival rate of the stem cells is high, and the preparation and preservation methods are simple and inexpensive.

Description

technical field [0001] The invention relates to the technical field of cell preservation, in particular to a umbilical cord blood hematopoietic stem cell preservation solution and a preparation method thereof. Background technique [0002] Umbilical cord blood is the blood remaining in the placenta and umbilical cord after the fetus is delivered and the umbilical cord is ligated and severed. Umbilical cord blood contains a large number of hematopoietic stem cells that can rebuild the human hematopoietic and immune system, and can be used for hematopoietic stem cell transplantation to treat various diseases. At the same time, its The collection is convenient, the source is sufficient, and there is no adverse effect on mothers and infants, so it is widely used. Usually, umbilical cord blood hematopoietic stem cells need to be preserved for further culture or clinical application after a period of time. Traditional umbilical cord blood preservation usually adopts the slow free...

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Problems solved by technology

[0003] Chinese patent CN201510120370.3 discloses a method for separating and building a bank of umbilical cord blood hematopoietic stem cells. In this method, dextran and dimethyl sulfoxide are mixed at a ratio of 1:1 in the cryopreservation process, and pre-cooled at 4°C for 15 minutes as a freezer. Store the solution, and then use the program cooling device to cool down and freeze it in liquid nitrogen. This method is simple and convenient, with low separation cost and high separation efficiency, and avoids microbial contamination. However, the dimethylformamide contained in the freezing solution used in this method Sulfoxide has certain toxicity to cells. Improper cryopreservation and resuscitation may cause serious damage to cells.

However, the cryopreservation solution contains fetal bovine serum, and usually animal-derived serum has the risk of being infected with pathogens such as viruses and bacteria. At the same time, when the stem cells are resuscitated and used in clinical applications, some proteins may cause immune rejection of heterologous proteins, and then Affect the patient's health

[0005] Aiming at the problems of toxicity, risk, and complicated and expensive preservation methods in the umbilical cord blood hematopoietic stem cell preservation solution, and easy damage to the cells, a preservation solution and its preparation method should be found so that the preserved Stem cells have less cell damage after resuscitation, and the survival rate of stem cells is higher. At the same time, the preparation and storage methods are relatively simple and cheap

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