Stem cell gel preparation for maintaining activity of cryopreserved cells and application thereof

A technology of gel preparation and cell activity, which is applied in the above-mentioned field of stem cell gel preparation, stem cell gel preparation, gel preparation; it can solve the problems of insufficient stability and cell viability, and achieve a simple and fast thawing method, Protect cells and DNA from damage, good biocompatibility

Active Publication Date: 2017-03-29
北京汉氏干细胞科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the preparation has a good therapeutic effect, it still has certain deficiencies in terms of stability and cell viability.

Method used

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  • Stem cell gel preparation for maintaining activity of cryopreserved cells and application thereof
  • Stem cell gel preparation for maintaining activity of cryopreserved cells and application thereof
  • Stem cell gel preparation for maintaining activity of cryopreserved cells and application thereof

Examples

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Effect test

preparation example Construction

[0035] Preparation of cell suspension: use DMEM medium (complete medium) containing 10% fetal bovine serum at 37°C, 5% CO 2 The mesenchymal stem cells were routinely cultured under the above conditions. When the cell confluence reached about 90%, they were digested with 0.25% trypsin, and then stopped with the above-mentioned complete medium, and the obtained cell suspension was centrifuged, and the supernatant was discarded. , and then suspended the cell pellet with serum-free DMEM medium, and adjusted the content of mesenchymal stem cells in the cell suspension to 0.4×10 6 ～ 2.5×10 7 per milliliter.

[0036] Calculation of cell survival rate: cell survival rate (%)=total number of living cells / (total number of living cells+total number of dead cells)×100%

[0037] Calculation of cell yield: cell yield (%) = total number of viable cells at the detection point / total total number of initial viable cells × 100%

Embodiment 1

[0038] [Example 1]: screening of sodium alginate

[0039]Prepare 200 mesh, 100 mesh and 50 mesh sodium alginate powders into 1%, 2%, and 3% sodium alginate solutions respectively, add 3% dimethyl sulfoxide, 5% propylene glycol, and then add cryopreserved and resuscitated The mesenchymal stem cells were mixed, placed in a 4-degree refrigerator after mixing evenly, and the cell viability and cell yield were compared at 0, 1, and 6 hours. As can be seen from the experimental results, 200-mesh sodium alginate is the most suitable gel material for cell survival ( figure 1 ).

Embodiment 2

[0040] [Example 2]: Screening and optimization of gel preparation formula

[0041] In order to screen the best gel formula for maintaining cell viability, the components in the formula are screened one by one to select the best formula:

[0042] (1) Screening of sodium alginate gel concentration

[0043] Add 200-mesh sodium alginate powder into the phosphate solution. The composition of the phosphate solution is 0.02% potassium chloride, 0.0047% magnesium chloride, 0.1158% disodium hydrogen phosphate, 0.020% sodium dihydrogen phosphate, 0.8% sodium chloride and water for injection. Prepare 1%, 2%, 3%, 4% sodium alginate aqueous solution, observe the clarity and fluidity of the gel. The 1%, 2% and 3% gels had good clarity, and the 4% gel had insolubles. The fluidity of the gel decreased with the increase of the concentration of sodium alginate, and the fluidity of the 3% gel was the least, and the adhesion ability was the best. Based on the results of clarity and fluidity, 3...

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Abstract

The invention relates to a gel preparation for maintaining the activity of cryopreserved cells and a stem cell gel preparation containing stem cells. The gel preparation comprises the following components by the mass percentage: 1-3% of sodium alginate, 1-5% of dimethyl sulfoxide, 1-5% of propylene glycol, 1-5% of epigallocatechin gallate, 2-10% of dextran, 1-5% of human blood albumin, and water or a phosphate solution added to make the total amount be 100%. The gel preparation has the advantages of good biocompatibility and simple operation, can maintain high activity of cells, can be directly used for low temperature preservation, and has no need of traditional liquid nitrogen cryopreservation; the cell gel preparation after resuscitation can still maintain high activity of the cells and dryness of the cells, and can be applied for treatment of skin injury or mucosal injury.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a gel preparation for cryopreservation of cells; [0002] The present invention also relates to a stem cell gel preparation containing mesenchymal stem cells and the above-mentioned gel preparation; [0003] The present invention also relates to the application of the above-mentioned stem cell gel preparation. Background technique [0004] To be used as a permanent substitute, tissue engineering scaffold materials must be degradable biomaterials, the materials themselves should not be immunogenic to tissues, and the degradation products of the materials should have no toxicity or abnormal reactions to biological tissues. Although collagen has a good affinity for cells, it is often derived from animal tissues, which has a high risk of contamination by pathogenic microorganisms and is immunogenic. It is also expensive and has poor mechanical properties, so it needs to be used after cr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02A61K35/51A61K9/06A61P17/02
CPCA61K47/36A01N1/0226A61K9/0014A61K9/06A61K35/51A61K35/28A01N1/02
Inventor 耿洁梁璐韩忠朝
Owner 北京汉氏干细胞科技有限公司
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