Human mesenchymal stem cell preservation transportation liquid and application thereof

A technology of mesenchymal stem cells and transport fluid, applied in the field of human mesenchymal stem cell preservation and transport fluid, can solve the problems of cell mass embolism of blood vessels, risk of patient infusion, death and other problems

Active Publication Date: 2017-03-29
杭州哈佛赛尔干细胞技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] An important issue in stem cell therapy is how to solve the problem of stem cell transportation, because there is a process between the preparation and use of stem cells, and this problem leads to three key technical problems: one is how to ensure cell viability during the transportation process, and how to At present, human mesenchymal stem cells can only be applied in a small area in the city and extended to the whole country, and even neighboring countries; second, the means of transportation and materials require safety and stability; third, mesenchymal stem cells are adherent cells, and after being prepared into a suspension It is easy to gather into clusters, and blood transfusion of mesenchymal stem cells is likely to cause cell clusters to embolize blood vessels, causing chest tightness, dizziness, and severe death
Moreover, after a few hours of storage, a large number of mesenchymal stem cells will aggregate into clusters, which will bring danger to patients during infusion.

Method used

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  • Human mesenchymal stem cell preservation transportation liquid and application thereof
  • Human mesenchymal stem cell preservation transportation liquid and application thereof
  • Human mesenchymal stem cell preservation transportation liquid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The effect of serum concentration of human AB on cell adhesion

[0025] The umbilical cord mesenchymal stem cells were prepared by the tissue block adherence method, and after three passages, the cells were collected by centrifugation, and the supernatant was removed. Prepare 5 groups of stem cell storage and transportation solutions, all of which contain 5.26mg / ml sodium chloride, 5.02mg / ml sodium gluconate, 3.68mg / ml sodium acetate, 2.5mg / ml ginsenoside Rb1, 10ng / ml transferrin , 10ng / ml reduced glutathione, 0.1mmol / l glutamine, wherein 2v / v% human AB serum in group A; group B: 5v / v% human AB serum; group C: 20v / v% human AB serum; D group: 50v / v% human AB serum; E group: 20v / v% human serum albumin. Except for the difference in serum concentration, the other components of each group were the same. Dilute the above-prepared umbilical cord mesenchymal stem cells into 5х10 7 / ml. 1ml of each group was stored statically in a 4°C incubator for 24 hours, then transferred...

Embodiment 2

[0027] Effects of transferrin and reduced glutathione on cellular ROS levels

[0028] The umbilical cord mesenchymal stem cells were prepared by the tissue block adherence method, and after three passages, the cells were collected by centrifugation, and the supernatant was removed. Prepare 2 groups of stem cell storage and transportation solutions, both of which contain 5.26mg / ml sodium chloride, 5.02mg / ml sodium gluconate, 3.68mg / ml sodium acetate, 2v / v% human AB serum, 2.5mg / ml ginsenoside Rb1, 0.1mmol / l glutamine, where A: without transferrin and reduced glutathione; B: with 10ng / ml transferrin and 10ng / ml reduced glutathione. Except for transferrin and reduced glutathione, the other components of the two groups were the same. Dilute the above-prepared umbilical cord mesenchymal stem cells into 5х10 7 / ml. 1ml of each group was statically stored in a 4°C incubator for 24 hours, centrifuged to remove the preservation solution, and the cells were washed three times with PBS...

Embodiment 3

[0030] Ginsenoside Rb1 can effectively maintain cell viability and metabolism

[0031] Conventional MTT method and PI staining method can only measure apoptotic and necrotic cells and non-apoptotic cells. In fact, many cells are irreversible when the environmental conditions are not suitable and their own regulation mechanism is not enough to regulate. It will gradually move from the normal state to apoptosis, and there is still a process from the beginning of apoptosis to necrosis, generally ranging from a few hours to a day or two depending on the treatment conditions. During this process, the cell membrane is still intact, PI cannot be stained, and the cell viability will change, but it can still maintain a certain metabolic capacity. However, at this time, the synthesis ability and content of ATP in the cell will change significantly. Therefore, ATP The content change of can best reflect the change of cell metabolism state.

[0032] The umbilical cord mesenchymal stem cel...

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Abstract

The invention discloses human mesenchymal stem cell preservation transportation liquid which is prepared from 1 to 15 mg/ml of sodium chloride, 1 to 15 mg/ml of sodium gluconate, 1 to 10 mg/ml of sodium acetate, 0.1 to 9.9 v/v% of human AB serum, 0.1 to 25 mg/ml of ginsenoside Rb1, 1 to 100 ng/ml of transferring, 1 to 100 ng/ml of reduced glutathione and 0.01 to 1 mmol/l of glutamine. The invention further discloses application of the liquid in preservation and transportation of human mesenchymal stem cells. The human mesenchymal stem cell preservation transportation liquid can reduce the clustering phenomenon of the human mesenchymal stem cells in a transportation process and keep high attachment rate and activity, so that the human mesenchymal stem cell preservation transportation liquid can be conveyed to all parts of the country and surrounding countries through modern vehicles such as an airplane, and the clinical use range of the human mesenchymal stem cells is greatly enlarged.

Description

technical field [0001] The invention relates to the technical field of tissue preservation, in particular to a human mesenchymal stem cell preservation transport solution and its application. Background technique [0002] Human mesenchymal stem cells (MSCs) are cells that can regenerate itself when an organ or tissue in our body is damaged. Using stem cell technology can regenerate a new, normal or even younger cell, tissue or organ. Therefore, people can use their own or others' stem cells and stem cell-derived tissues and organs to replace diseased or aging tissues and organs, and can be widely used to treat various chronic diseases that are difficult to cure by traditional medical methods, such as leukemia, Alzheimer's disease, Parkinson's disease, etc. Disease, diabetes, stroke, chronic diseases of unknown cause such as pharyngitis and a series of diseases that are currently incurable. Theoretically speaking, the application of stem cell technology can treat various dis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0226
Inventor 范汉东姜明明屈彩云
Owner 杭州哈佛赛尔干细胞技术有限公司
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