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Preserving solution for maintaining activity of mesenchymal stem cells in transportation and preparation method thereof

A technique of mesenchymal stem cells and preservation solution, which is applied in the field of preservation solution for maintaining the activity of mesenchymal stem cells and its preparation, can solve the problems of limiting the application scope of MSCs, affecting the therapeutic effect of MSCs, and limited effect, and achieving the expansion of the geographical scope of clinical use, The effect of maintaining vitality and good protection

Active Publication Date: 2017-11-24
COBAXER BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effects of these protective agents are limited, and the viability maintenance time is generally about 12 hours. After long-term storage (24 hours), the cell viability will drop below 90%, which limits the application range of MSCs to a certain extent, and also affects MSCs. The therapeutic effect of MSCs

Method used

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  • Preserving solution for maintaining activity of mesenchymal stem cells in transportation and preparation method thereof
  • Preserving solution for maintaining activity of mesenchymal stem cells in transportation and preparation method thereof
  • Preserving solution for maintaining activity of mesenchymal stem cells in transportation and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 The protective effect of compound amino acids on mesenchymal stem cells

[0028] In order to compare the protective effects of compound amino acids and glucose on the viability of mesenchymal stem cells, the mesenchymal stem cell transport preservation solution was divided into 3 groups of different formulations in this example.

[0029] Group 1: without cell viability protection agent: Mix 75ml of compound electrolyte injection, 25ml of stock solution concentration of 20% human albumin (final concentration of 0.05g / ml).

[0030] Group 2: only include glucose: 72ml of compound electrolyte injection, 20% human serum albumin 25ml (final concentration of 0.05g / ml) and 3ml of glucose injection (final concentration of 0.05g / ml) with stock solution concentration 1.5mg / ml) and mix well.

[0031] Group 3: only containing compound amino acids: compound amino acid injection (18AA) with 74ml of compound electrolyte injection, 25ml of human serum albumin (final concentra...

Embodiment 2

[0037] Example 2 The protective effect of melatonin and rapamycin on mesenchymal stem cells

[0038] In order to further optimize the formulation of the transport preservation solution, in this example, on the basis of adding compound amino acids, the cell viability protective agents melatonin and rapamycin were added, and the effects of melatonin and rapamycin on mesenchymal stem cells were determined. Vitality protection. In this example, the mesenchymal stem cell transport preservation solution was divided into 3 groups.

[0039] Group 4: Containing compound amino acid and melatonin: compound amino acid injection with 73ml compound electrolyte injection, 20% stock solution concentration of human albumin 25ml (final concentration 0.05g / ml), stock solution concentration 0.09g / ml (18AA) 1ml (final concentration: 0.9mg / ml), stock solution concentration: 0.115mg / ml melatonin solution 1ml (final concentration: 1.15μg / ml) and mix well.

[0040] Group 5: Containing compound amino...

Embodiment 3

[0048] Example 3 The protective effect of different preservation solution formulations on mesenchymal stem cells

[0049] In order to compare the transport preservation solution in the present invention with the mesenchymal stem cell preservation solution and injection solution in other patents, this embodiment selected two reported formulas and the formula of the present invention (Example 2, Group 6) for cell viability Determination test.

[0050] Patent A formula: normal saline, human serum albumin (5%), low molecular weight heparin calcium (75AXaIU / mL)

[0051] Patent B formula: normal saline, autologous platelet lysate (4%), vitamin C injection (1%)

[0052] Formula of the present invention: compound electrolyte injection, human serum albumin (5%), compound amino acid injection (0.9mg / ml), melatonin (1.15 μ g / mL), rapamycin (100nM)

[0053] The transport solution of the above-mentioned groups of formulas was filtered and sterilized with a filter with a pore size of 0.2 ...

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Abstract

The invention discloses a preserving solution capable of effectively maintaining the activity of mesenchymal stem cells in transportation and a preparation method thereof. The preserving solution is composed of a solution medium and a cell activity protecting agent, wherein the solution medium is a compound electrolyte injection with the addition final concentration of 0.05g / ml of human serum albumin; the cell activity protecting agent comprises compound amino acid, melatonin and rapamycin with the final concentrations of 0.4-1.6mg / ml, 0.23-2.3mu g / ml and 20-200nM respectively; and the compound amino acid can provide energy for the mesenchymal stem cells, and the melatonin and rapamycin can activate the autophagy effect of the cells and promote the viability of the mesenchymal stem cells. The preserving solution for transportation provided by the invention can effectively maintain the activity of the mesenchymal stem cells in 24 hours and guarantee the stability of the stem cells in long-distance transportation.

Description

technical field [0001] The invention relates to the technical field of stem cell preservation solution, in particular to a preservation solution for maintaining the activity of mesenchymal stem cells and a preparation method thereof. Background technique [0002] Mesenchymal stem cells (Mesenchymal Stem Cells, MSCs) are derived from the mesoderm in the early stage of development. They are a type of primitive cells with self-replication and multi-directional differentiation potential. , fat, etc.; under specific conditions, it can differentiate into a variety of tissue-specific cells, such as osteoblasts, chondrocytes, fat cells, nerve cells, cardiomyocytes, etc. MSCs have immune regulation and tissue repair capabilities, and act on target cells through paracrine effects to achieve the purpose of treating graft-versus-host disease, inflammatory disease, bone disease, heart disease, and nervous system disease. [0003] At present, allogeneic general-purpose cell products are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0226
Inventor 左凤琼陈勇军幸倚帆程威谭小虎
Owner COBAXER BIOTECH
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