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Low-temperature storage liquid for cells, sodium alginate gel, cell gel, preparation method, use method and application thereof

A low-temperature preservation and sodium alginate technology, which is applied in the field of cell preservation and regeneration and repair technology, can solve problems such as affecting experimental or clinical effects, rapid decline in cell activity, and difficulty in activation, so as to facilitate subsequent application, maintain cell activity, The effect of maintaining stability

Pending Publication Date: 2019-01-18
CHENGDU YUANSHAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this storage method has the following problems: cell activity decreases rapidly during transportation, the survival rate of cells after long-term transportation is very low, and a large number of cells die, which directly affects subsequent experiments or clinical effects; cell recovery requires high operation and is not easy activation; in addition, most cryopreservation solutions need to add DMSO and other substances that are toxic to tissue cells, which greatly limits the clinical application of cell products
[0007] "A Stem Cell Gel for Maintaining the Activity of Frozen Cells and Its Application" (Chinese Invention Patent Publication No. CN106538512A), prepared by using sodium alginate and dimethyl sulfoxide, etc. The shelf life of stem cells can be extended to 18 months under cryopreservation at ℃, but this kind of gel still needs to be stored at a low temperature of -80 ℃ to -60 ℃, and dimethyl sulfoxide is added. It is necessary to recover the gel (the cells in it), which is not convenient for the storage, transportation and clinical application of the gel

Method used

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  • Low-temperature storage liquid for cells, sodium alginate gel, cell gel, preparation method, use method and application thereof
  • Low-temperature storage liquid for cells, sodium alginate gel, cell gel, preparation method, use method and application thereof
  • Low-temperature storage liquid for cells, sodium alginate gel, cell gel, preparation method, use method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The purpose of this embodiment is to screen the viscosity of sodium alginate.

[0045] Sodium alginate powder commonly used in the market is divided into three types: high viscosity (300-500CPS), medium viscosity (100-300 CPS) and low viscosity (50-100CPS). Mix high-viscosity, medium-viscosity and low-viscosity sodium alginate powders into 5% sodium alginate solutions with physiological saline, sterilize under high pressure steam at 121°C for 25 minutes, add 1% calcium chloride solution for cross-linking, and compare the three Clarity, viscosity and hardness of the gel formed by sodium alginate. Judging method: ++++ means good, +++ means medium, ++ means average, + means poor, and the follow-up screening experiment is the same. The results are shown in Table 1.

[0046] Table 1

[0047]

[0048] It can be seen from Table 1 that the high-viscosity sodium alginate cross-linked gel has the best viscosity and hardness, medium transparency, and moderate overall ...

Embodiment 2

[0050] The purpose of this example is to screen the concentration of sodium alginate.

[0051] Add high-viscosity sodium alginate powder into physiological saline to prepare 1%, 2%, 3%, 4%, 5% sodium alginate solution, sterilize by high-pressure steam at 121°C for 25 minutes, add 1% calcium chloride solution for cross-linking , to compare the transparency, viscosity and hardness of gels cross-linked with different concentrations of sodium alginate solutions. The results are shown in Table 2.

[0052] Table 2

[0053]

[0054] It can be seen from Table 2 that the viscosity of the gel formed by the cross-linking of 1% sodium alginate solution is low, and the degree of cross-linking is not enough; the hardness of sodium alginate above 4% is too high, and the cross-linking is too high; 2% and 3% sodium alginate solution The texture of the gel formed by cross-linking is moderate, so it can be seen that the optimum concentration of sodium alginate is 2% to 3%.

Embodiment 3

[0056] The purpose of this embodiment is to screen the concentration of calcium chloride.

[0057] High-viscosity sodium alginate powder was used to prepare a 3% sodium alginate solution, and sterilized by high-pressure steam at 121° C. for 25 minutes. The optimal osmotic pressure range of the solution during cell culture is 290mmol / L-310mmol / L, the calcium ion concentration is 1.8-2.8mmol / L, and the chloride ion concentration is 130-160mmol / L. In order to reduce the influence of the calcium chloride used for cross-linking on the osmotic pressure as much as possible, an equal volume of calcium chloride solution with a concentration of 0.2%, 0.4%, 0.6%, 0.8% and 1.0% was set for cross-linking according to the calculation. Determine the optimum calcium chloride solution concentration for crosslinking. The cross-linking situation of different calcium chloride concentrations is shown in Table 3.

[0058] table 3

[0059] Calcium chloride concentration

[0060] It can ...

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Abstract

The invention belongs to the technical field of storage, regeneration and repair of cells and discloses low-temperature storage liquid for cells. The low-temperature storage liquid is mainly preparedfrom the following components in percent by mass: 0.8%-1.5% of mannitol, 2%-3% of human serum albumin, 3%-7% of low-molecular dextran, 0.1-0.3% of glutathione, 0.5%-2% of epigallocatechin gallate, 3%-5% of adenosine, 0.5%-1% of glucose, 0.8% of NaCl, 0.2% of KCl, 0.1% of CaCl2 and the balance of water. The invention also provides sodium alginate gel, which contains sodium alginate, calcium chloride, hyaluronic acid and the low-temperature storage liquid for the cells and the like, and also provides cell gel and a preparation method thereof as well as application of the low-temperature storageliquid for the cells, the sodium alginate gel and the cell gel. The invention has the advantages that the low-temperature storage liquid for the cells can maintain cell activity for longer time at temperature of 2-8 DEG C, the sodium alginate gel can be used as temporary short-term storage materials of cells from various sources and brackets of cell tissue engineering, and the cell gel can be usedas a medical wet wound dressing and can be used for repairing soft tissues, refractory-healing wound skins and mucosal wound injuries.

Description

Technical field [0001] The invention relates to the technical field of cell preservation and regeneration and repair, specifically to a cell cryopreservation solution, a sodium alginate gel rich in three-dimensional pores prepared by using the cell cryopreservation solution, and also relates to a sodium alginate gel prepared by using the sodium alginate gel. Cell gel, and the present invention also relates to a preparation method of the cell gel and a method for isolating and activating cells therein. In addition, the invention also relates to a cell cryopreservation solution, sodium alginate gel and cell gel applications. Background technique [0002] Cells are widely used in scientific research and clinical practice, among which cell preservation and transportation are important parts of cell applications. At present, cell preservation generally involves storing cells at a low temperature of -196°C in liquid nitrogen, which can temporarily remove the cells from the growth ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02C12N5/0775A61L27/52A61L27/20A61L27/22A61L27/38A61L26/00
CPCA01N1/021A01N1/0221A61L26/0023A61L26/0047A61L26/008A61L27/20A61L27/227A61L27/3834A61L27/52C12N5/0662C12N2509/00C08L5/04C08L5/08C08L89/00
Inventor 杨远袁祥韩飞张中伟潘红伟
Owner CHENGDU YUANSHAN BIOTECH CO LTD
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