Cell cryopreservation method

Abstract

The invention relates to a cell cryopreservation method, belonging to the technical field of cell freezing. The cell cryopreservation method comprises the following steps: before collecting cells, adding saccharides into a cell culture solution until a final concentration is 50-400 mM, and after collecting the cells, resuspending the cells by using a sugar solution containing 50-250 mM of saccharides and conducting cryopreserving, wherein the saccharides comprise glucose, cane sugar and raffinose. The method disclosed by the invention is safe and efficient, can realize long-term low-temperature high-quality preservation of the cells, does not need to use components such as DMSO, glycerol and the like which are toxic to the cells, and is beneficial to development of cell drugs.

Description

technical field [0001] The invention relates to the technical field of cell freezing, in particular to a cell freezing method. Background technique [0002] Cell therapy involves stem cell therapy and immune cell therapy for various diseases including tumors. In recent years, cell therapy has become a frontier exploration field of life science and medicine. Innovative technology research and clinical research of cell therapy have contributed to the development of the cell therapy industry. A good foundation has been laid. The industrial chain of the cell therapy industry includes cell storage, cell medical technology, and cell drug development; the corresponding primary stem cells or immune precursor cells are extracted from umbilical cord, placenta, bone marrow and other tissues or umbilical cord blood and peripheral blood in a certain way. Cells as seed cells need to be stored for later use; seed cells can be further developed into medical technology or new cell drugs thr...

AI Technical Summary

Problems solved by technology

[0003] In the deep cryopreservation of cells, the traditional method is to collect target cells and use cryoprotectants for cryopreservation. The cryoprotectants used are divided into osmotic protective agents and non-osmotic protective agents. The osmotic protective agents include Organic solvents such as dimethyl sulfoxide (DMSO), glycerin, ethylene glycol, and propylene glycol, and non-permeable protective agents are mainly macromolecular substances, including dextran, hydroxyethyl starch, polyvinylpyrrolidone, albumin, and polyvinylpyrrolidone. Sucrose, etc., the preservation effect of different cell protectants on cells is very different, and multiple protectants need to be used in combination. Toxic and side effects, as well as risks such as potential allergic reactions to macromolecular substances such as dextran and albumin, even excipient-level DMSO also has side effects such as vascular stimulation and oral mucosal stimulation. Damaged, DMSO and macromolecular extracellular protective agents will increase the metabolic burden of the patient's liver, kidneys and other organs

[0004] On the other hand, the use of traditional protective agents containing hyperosmotic substances such as DMSO and glycerin also has great limitations on the large-scale preparation of cell drugs. Cells cannot survive in a hyperosmotic environment for more than 1 to 2 hours, even if there The cells will be greatly affected in terms of activity and function. Secondly, the higher the temperature of the osmotic protective agent DMSO, glycerin, the greater the toxicity to the cells, which requires strict control of the cell drug preparation environment. Low temperature conditions increase the difficulty of the preparation process, which is also the key and urgent technical difficulty that currently limits the development and application of cell drug processes

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