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Hematopoietic stem cell cryopreservation solution and cryopreservation method

A technology of hematopoietic stem cells and cryopreservation methods, applied in the preservation, application, and animal husbandry of human or animal bodies, which can solve the problems of poor safety and achieve the effects of maintaining stability, avoiding water crystallization, and changing permeability

Inactive Publication Date: 2021-05-28
赵丽歌
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Animal serum and non-clinical application grade reagents will bring many unknown risks and poor safety

Method used

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  • Hematopoietic stem cell cryopreservation solution and cryopreservation method

Examples

Experimental program
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Effect test

Embodiment 1

[0022] A cryopreservation solution for hematopoietic stem cells, including basal medium RPMI-1640 and additives, the ingredients of the additives in terms of final concentration include: rice protein 10 μg / mL, sulbutiamine 5 μg / mL, coumarin 8 μg / mL, carbamic acid Ethyl ester 12μg / mL, mannitol 25μg / mL, vitamin C 12ng / mL, naringin 15ng / mL.

[0023] A cryopreservation method for hematopoietic stem cells, comprising the following steps:

[0024] (1) Take the basal medium RPMI-1640 and add rice protein, sulbutiamine, coumarin, ethyl carbamate, mannitol, vitamins, and naringin, and stir evenly at 45°C to obtain a cryopreservation solution;

[0025] (2) Add hematopoietic stem cells to be preserved into the cryopreservation solution of step (1), and the concentration of hematopoietic stem cells in the cryopreservation solution is 3×10 7 pcs / mL, divided into 2mL cryovials, each containing 1.5mL, cooled the cryovials at a rate of 1°C / min, cooled to -20°C, and then put them into -80°C f...

Embodiment 2

[0027] A hematopoietic stem cell cryopreservation solution, including basal medium RPMI-1640 and additives, the ingredients of which are calculated by final concentration: rice protein 8 μg / mL, sulbutiamine 2 μg / mL, coumarin 5 μg / mL, carbamic acid Ethyl ester 10 μg / mL, mannitol 20 μg / mL, vitamin C 10 ng / mL, naringin 11 ng / mL.

[0028] A cryopreservation method for hematopoietic stem cells, comprising the following steps:

[0029] (1) Take the basal medium RPMI-1640 and add rice protein, sulbutiamine, coumarin, ethyl carbamate, mannitol, vitamins, and naringin, and stir evenly at 40°C to obtain a cryopreservation solution;

[0030] (2) Add hematopoietic stem cells to be preserved into the cryopreservation solution of step (1), the concentration of hematopoietic stem cells in the cryopreservation solution is 1×10 7 1 / mL, put into 2mL cryovials, each tube contains 1.5mL, cool the cryovials at a rate of 2°C / min, cool down to -25°C, and put them into -80°C for freezing.

Embodiment 3

[0032] A hematopoietic stem cell cryopreservation solution, including basal medium RPMI-1640 and additives. The ingredients of the additives include: rice protein 12 μg / mL, sulbutiamine 8 μg / mL, coumarin 10 μg / mL, carbamic acid Ethyl ester 15μg / mL, mannitol 30μg / mL, vitamin C 15ng / mL, naringin 18ng / mL.

[0033] A cryopreservation method for hematopoietic stem cells, comprising the following steps:

[0034] (1) Take the basal medium RPMI-1640 and add rice protein, sulbutiamine, coumarin, ethyl carbamate, mannitol, vitamins, and naringin, and stir evenly at 50°C to obtain a cryopreservation solution;

[0035] (2) Add hematopoietic stem cells to be preserved into the cryopreservation solution of step (1), the concentration of hematopoietic stem cells in the cryopreservation solution is 5×10 7 1 / mL, divided into 2mL cryovials, each containing 1.5mL, cooled the cryovials at a rate of 2°C / min, cooled to -30°C, and then put them into -80°C for cryopreservation.

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Abstract

The invention discloses a hematopoietic stem cell cryopreservation solution, which comprises a basic culture medium and an additive, and the additive comprises the following components by final concentration: 8-12 [mu] g / mL of rice protein, 2-8 [mu] g / mL of sulbrutiamine, 5-10 [mu] g / mL of coumarin, 10-15 [mu] g / mL of ethyl carbamate, 20-30 [mu] g / mL of mannitol, 10-15 ng / mL of vitamin C, and 11-18 ng / mL of naringin. According to the hematopoietic stem cell cryopreservation solution, DMSO is not added, through the synergistic effect of rice protein, sulbrutiamine, coumarin, ethyl carbamate, mannitol, vitamin C and naringin, the survival rate of hematopoietic stem cells under the preservation condition of-80 DEG C is increased, the cells are easily separated from the hematopoietic stem cells after resuscitation, cytotoxicity is avoided, and the clinical application safety of the hematopoietic stem cells is improved. The invention also provides a cryopreservation method of the hematopoietic stem cells, the cryopreservation of the hematopoietic stem cells is realized at -80 DEG C by adopting the cryopreservation liquid provided by the invention, the operation is simple and convenient, and the hematopoietic stem cell cryopreservation method is easy to popularize and apply.

Description

technical field [0001] The invention relates to a cryopreservation solution, in particular to a hematopoietic stem cell cryopreservation solution and a cryopreservation method. Background technique [0002] Hematopoietic stem cells (HSCs) are adult stem cells in the blood system. They are a heterogeneous population with the ability of long-term self-renewal and the potential to differentiate into various mature blood cells. It is a type of adult stem cell with the longest and most in-depth research history, and it has important guiding significance for the study of various stem cells, including tumor stem cells. Mature cells in the blood system have a very short lifespan. Therefore, in a person's life, hematopoietic stem cells need to replenish various mature cell components of the blood system in a timely manner according to the physiological needs of the body. At the same time, under stress conditions such as injury and inflammation, hematopoietic stem cells also play a r...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226A01N1/0284
Inventor 赵丽歌张少苹
Owner 赵丽歌
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