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Cryoprotectant for mesenchymal stem cells, and preparation method of cryoprotectant

A technology for cryoprotective solution and quality stem cells, applied in the field of stem cell cryopreservation, can solve problems such as adverse effects on cell growth, cytotoxicity, etc., and achieve the effects of improving preservation vitality, avoiding stress-induced damage, and improving cell survival rate.

Active Publication Date: 2019-10-01
CENTURY BIOSTRENGTH BEIJING PTY LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But DMSO is easy to cause serious toxicity to cells; the research results show that when the DMSO concentration in the culture medium is 10%, the cell growth inhibition rate is nearly 100%; the inhibition rate is 35% when the ‰ concentration is 0.04‰ Cell growth can also be adversely affected

Method used

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  • Cryoprotectant for mesenchymal stem cells, and preparation method of cryoprotectant
  • Cryoprotectant for mesenchymal stem cells, and preparation method of cryoprotectant
  • Cryoprotectant for mesenchymal stem cells, and preparation method of cryoprotectant

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037]

Embodiment 2

[0039]

[0040] Wherein, the succinylated gelatin microspheres consist of the following components in parts by weight: 0.15 parts by weight of succinic anhydride; 2.5 parts by gelatin; and 1.5 parts by weight.

[0041] Described preparation method comprises:

[0042] 1) Preparation of succinylated gelatin microspheres

[0043] Dissolving the gelatin by weight in water, adding an appropriate amount of sodium chloride to the dissolved gelatin solution to adjust the pH value to 8-10; adding the succinic anhydride in the weight part for acylation reaction to obtain succinylated gelatin solution;

[0044] Dissolving the cichoric acid in the weight portion in the succinic acid gelatin solution, and dissolving in 4% (w / v) liquid paraffin mixed with an emulsifier, stirring evenly, stirring at a speed of 200 r / min for 25 min to form an emulsion; Add isopropanol and stir, and filter with suction to obtain a filter cake; add 5 times the volume of the filter cake to an isopropanol so...

Embodiment 3

[0049]

[0050] Wherein, the succinylated gelatin microspheres consist of the following components in parts by weight: 1.34 succinic anhydride; 4.79 gelatin; 2.24 cichoric acid.

[0051] Described preparation method comprises:

[0052] 1) Preparation of succinylated gelatin microspheres

[0053] Dissolving the gelatin by weight in water, adding an appropriate amount of sodium chloride to the dissolved gelatin solution to adjust the pH value to 8-10; adding the succinic anhydride in the weight part for acylation reaction to obtain succinylated gelatin solution;

[0054] Dissolving the cichoric acid in the weight portion in the succinic gelatin solution, and dissolving in 4% (w / v) liquid paraffin mixed with an emulsifier, stirring evenly, stirring at a speed of 350 r / min for 25 min to form an emulsion; Add isopropanol and stir, and filter with suction to obtain a filter cake; add 5 times the volume of the filter cake to an isopropanol solution containing a crosslinking curi...

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Abstract

The invention provides a cryoprotectant for mesenchymal stem cells. The cryoprotectant is mainly prepared from the following components in parts by weight: 60-80 parts of physiological saline, 2.2-5.87 parts of succinylated gelatin microspheres, 1.40-3.11 parts of L-histidine hydrochloride and 0.3-0.8 part of mannitol. The cryoprotectant can protect the mesenchymal stem cells for a long time, andcan maintain cell viability.

Description

technical field [0001] The invention belongs to the technical field of stem cell cryopreservation, and in particular relates to a cryoprotective solution for mesenchymal stem cells and a preparation method thereof. Background technique [0002] Mesenchymal stem cells are a kind of stem cells derived from mesoderm with multi-lineage differentiation ability, which widely exist in various tissues of the body, can be cultured and expanded in vitro, and can differentiate into cardiomyocytes, nerve cells and fat under specific conditions cells etc. With the in-depth research on the biological characteristics and functions of mesenchymal stem cells by scholars at home and abroad, mesenchymal stem cells can be isolated from various tissues. [0003] In the prior art, physiological saline or cell culture medium is usually used as the solution medium for clinical application of mesenchymal stem cells. If the human mesenchymal stem cells are isolated, cultured, and expanded, and imme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 董明珠李霞云潘新刘世红卢家堃
Owner CENTURY BIOSTRENGTH BEIJING PTY LTD
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