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Cell freezing medium, freezing resuscitation method and application thereof

A cryopreservation, cell suspension technology, applied in the field of clinical medicine, can solve problems such as cell damage, and achieve the effects of reducing damage, reducing cell damage, and improving cell activity

Active Publication Date: 2019-01-18
SHANGHAI CRYOWISE MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cell cryopreservation solution disclosed by it causes damage to the cells to a certain extent when the cells are preserved for a long time, and the cell survival rate needs to be further improved.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] The cell cryopreservation solution of this embodiment and its preparation method include: the preparation of the equilibrium buffer, the preparation of the acylated ε-polylysine solution, and the preparation of the equilibrium buffer and the acylated ε-polylysine solution The preparation of the main cell cryopreservation solution will be described in detail later.

[0108] 1. The preparation of the equilibrium buffer includes the following steps: add NaCl 100mmol, KCl 10mmol, MgCl to ultrapure water in sequence 2 13mmol, CaCl 2 0.25mmol, after all the above substances are completely dissolved, adjust the pH of the above solution to 7.0~7.6 with 10mol / L NaOH aqueous solution; then dilute to 1L with ultrapure water.

[0109] 2. Preparation steps of acylated ε-polylysine solution: Weigh 13g of succinic anhydride and pour it into 100mL of ε-polylysine aqueous solution with a mass concentration of 0.25g / mL, and put it into 50°C water for water bath , after 2 hours of water...

Embodiment 2

[0116] The cell cryopreservation solution of this embodiment and its preparation method include: the preparation of the equilibrium buffer, the preparation of the acylated ε-polylysine solution, and the preparation of the equilibrium buffer and the acylated ε-polylysine solution The preparation of the main cell cryopreservation solution will be described in detail later.

[0117] 1. The preparation of the equilibrating buffer comprises the following steps:

[0118] Step A: Add NaNO sequentially to ultrapure water 3 15mmol, KNO 3 10mmol, Mg(NO 3 ) 2 10mmol, Ca(NO 3 ) 2 0.5mmol, dissolve all the above substances completely;

[0119] Step B: Add 15mmol of mannitol, 1mmol of vitamin C, 3mmol of glutathione, 2mmol of N-acetyl-L-cysteine, 2mmol of tert-butylterephthalene, and 2 μmol of phenol, 20 mmol of histidine, 14 mmol of glutamic acid, 1 mmol of α-ketoglutarate, and 5 mmol of adenosine, until all substances are completely dissolved;

[0120] Step C: then adjust the pH o...

Embodiment 3

[0129] The cell cryopreservation solution of this embodiment and its preparation method include: the preparation of the equilibrium buffer, the preparation of the acylated ε-polylysine solution, and the preparation of the equilibrium buffer and the acylated ε-polylysine solution The preparation of the main cell cryopreservation solution will be described in detail later.

[0130] 1. The preparation of the equilibrating buffer comprises the following steps:

[0131] Step A1: Add the NaCl 100mmol, the KCl 10mmol, the MgCl to the ultrapure water successively 2 13mmol, the CaCl 2 0.25mmol, dissolve all the above substances completely;

[0132] Step B1: Add 10 mmol of mannitol, 0.5 mmol of vitamin C, 2 mmol of glutathione, 5 mmol of N-acetyl-L-cysteine, and 5 mmol of tert-butyl-p-phenylene in sequence. 5 μmol of diphenol, 24.5 mmol of histidine, 16.3 mmol of glutamic acid, 0.5 mmol of α-ketoglutaric acid, 3 mmol of adenosine and 15 g of human serum albumin, until all substances ...

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Abstract

The invention provides a cell freezing medium. The cell freezing medium comprises acylated epsilon polylysine, polyhydric alcohols and equilibration buffer. According to the cell freezing medium disclosed by the invention, the polyhydric alcohols and acylated epsilon polylysine replace toxic dimethyl sulfoxide so as to serve as cryoprotective agents, so that damage to cells in the freezing resuscitation process is reduced, and the cell survival rate is improved. The invention further provides a method for performing freezing resuscitation on cells by utilizing the cell freezing medium and application of the method for freezing stem cells and immune cells. The method is simple and feasible to operate and is suitable for large-scale clinical application of cell therapy, and great convenienceis brought to clinical application of the cell freezing technology.

Description

technical field [0001] The invention relates to the technical field of clinical medicine, in particular to a cell cryopreservation solution, a cell cryopreservation recovery method and applications thereof. Background technique [0002] Cell therapy plays an important role in various disease areas. Cell therapy is a treatment method that uses adult cells or stem cells to repair tissues or organs. Among them, stem cells and immune cells are the seed cells for disease treatment. [0003] Due to their potential for self-renewal and multidirectional differentiation, stem cells are the origin cells of the body and the original cells that form various tissues and organs of the human body. They have been widely used in scientific research and clinical treatment. With the development of cell replacement therapy, stem cell transplantation has become an important means of clinical treatment of certain diseases. At present, there are stem cell drug products on the market around the w...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N5/00C12N5/0775
CPCA01N1/0221C12N5/00C12N5/0662
Inventor 鄢和新杨秋蕊翟志敏
Owner SHANGHAI CRYOWISE MEDICAL TECH CO LTD
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