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Trehalose derivative as well as cryoprotectant and application thereof

A cryoprotectant, trehalose technology, applied in the application, preservation of human or animal body, animal husbandry, etc., can solve the problems of low cell permeability of trehalose, poor low temperature preservation effect, etc.

Inactive Publication Date: 2021-12-03
章毅 +7
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cell permeability of trehalose is low, resulting in poor cryopreservation effect

Method used

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  • Trehalose derivative as well as cryoprotectant and application thereof
  • Trehalose derivative as well as cryoprotectant and application thereof
  • Trehalose derivative as well as cryoprotectant and application thereof

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preparation example Construction

[0029] The preparation method of gallic acid grafted chitosan scaffold of the present invention is as follows:

[0030] First, 28 mmol of gallic acid and 2.8 mmol of EDC were dissolved in 40 mL of 70% ethanol, and 2.8 mmol of NHS was added to the solution. The resulting solution was stirred in an ice bath, then 1.5 g of chitosan dispersed in 110 mL of 70% ethanol was added after 1 hour. The solution was stirred for a further 30 minutes in an ice bath and finally at room temperature for 24 hours. The precipitate was collected by filtration and washed with ethanol. After dialysis against deionized water for 3 days to remove possible residual reagents, gallic acid-grafted chitosan was obtained by vacuum freeze-drying. Porous gallic acid grafted chitosan scaffolds were prepared by using the physical properties of gallic acid grafted chitosan dissolved in acidic solution and insoluble in alkaline condition. First, 1 g of gallic acid-grafted chitosan was dissolved in 50 ml of 0.1...

Embodiment 1

[0042] For the survival rate of mesenchymal stem cells in tissue engineered structures, see figure 1 shown.

[0043] For the proliferative ability of mesenchymal stem cells in tissue engineered structures, see image 3 shown.

[0044] For the colony-forming ability of mesenchymal stem cells in tissue engineering structures, see Figure 4 As shown, the three differentiation abilities of mesenchymal stem cells in tissue engineering structures can be found in Figure 5 shown.

Embodiment 2

[0046] The tissue engineered constructs were cryopreserved in fetal bovine serum (90% FBS) cryovials containing 10% DMSO. The cryopreservation tubes were then stored at 4°C for 30 minutes, at -20°C for 60 minutes, at -80°C overnight, and finally in liquid nitrogen for 30 days.

[0047] For the survival rate of mesenchymal stem cells in tissue engineering structures after cryopreservation, see figure 1 shown. For the survival rate of mesenchymal stem cells in tissue engineering structures after cryopreservation, see figure 2 shown.

[0048] For the proliferative ability of mesenchymal stem cells in tissue engineering structures after cryopreservation see image 3 As shown, the colony-forming ability of mesenchymal stem cells can be found in Figure 4 As shown, the three differentiation abilities of mesenchymal stem cells can be found in Figure 5 shown.

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PUM

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Abstract

The invention discloses a trehalose derivative which comprises trehalose and epichlorohydrin, and the trehalose and the epichlorohydrin are in covalent binding. The trehalose derivative disclosed by the invention is prepared into a cryoprotectant, is suitable for low-temperature cryopreservation of a tissue engineering structure (such as a scaffold) containing living cells, is used as a substitute of DMSO, reduces the use of DMSO, maintains the survival rate and the recovery rate of the cells at a relatively high level, and can enable frozen stem cells to continuously maintain dryness and differentiation potential after recovery.

Description

technical field [0001] The present invention relates to a polysaccharide derivative, in particular to a polysaccharide covalent derivative, which has the function of protecting cells in deep freezing of cells and serves as a substitute for DMSO. Background technique [0002] Tissue engineering and regenerative medicine are considered to be a promising approach to repair damaged tissues and maintain biological functions. Among the therapeutic methods of tissue engineering and regenerative medicine, cell-based therapy is one of the recent hotspots. These therapies have been focused on regenerating damaged tissue and delivering many beneficial cytokines. In particular, implanting stem cells in scaffolds allows stem cells to maintain their stemness. In previous studies, we established tissue engineering constructs based on gallic acid-grafted chitosan scaffolds and mesenchymal stem cells. In addition, the difficulty of scaffold fabrication is to encapsulate cells into scaffol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00A01N1/02
CPCC08B37/006A01N1/0221
Inventor 章毅伍婷胡肖希陈亮蔡海波
Owner 章毅
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