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Special normal-temperature preservation fluid for dog mesenchymal stem cell and application

A kind of stem cell storage technology at room temperature, applied in the field of bioengineering, can solve the problems of complex processing methods and high cost, and achieve the effects of low production cost, easy preparation, and prevention of death or clustering

Inactive Publication Date: 2018-04-17
FOSHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the disadvantages and deficiencies of the high cost and complicated treatment methods of the above-mentioned technologies, the present invention provides a preservation solution of mesenchymal stem cells that can be transported for a long time and long distance at room temperature, especially mesenchymal stem cells derived from dogs Stem cells are stored in this preservation solution, which is convenient for storing and transporting MSCs, and ensures the viability and stability of cells

Method used

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  • Special normal-temperature preservation fluid for dog mesenchymal stem cell and application
  • Special normal-temperature preservation fluid for dog mesenchymal stem cell and application
  • Special normal-temperature preservation fluid for dog mesenchymal stem cell and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: Culture of canine mesenchymal stem cells.

[0014] Taking canine adipose-derived mesenchymal stem cells as an example, about 1 cm of healthy dog ​​inguinal adipose tissue was collected on a sterile operating table. 2 In a 50ml centrifuge tube, wash the blood with normal saline, cut the tissue pieces into pieces with dissecting scissors, add 5ml of 1mg / ml type Ⅰ collagenase and place it in a water bath at 37°C for 1h, take it out and filter it with a 200-mesh sieve, the filtrate Centrifuge at 1200rmp for 5min to discard the supernatant, resuspend the pellet in DMEM culture solution containing 10% fetal bovine serum, 1% double antibody, and 1% L-glutamine, transfer it to a petri dish, and place it at 37°C in 5% CO 2 in the incubator. Change the medium for the first time after 24 hours to wash away non-adherent cells and impurities, and change the medium every 2-3 days thereafter. Observe the state of the cells every day, and pass passage when the adhesion re...

Embodiment 2

[0015] Embodiment 2: the preparation of special room temperature preservation solution.

[0016] Solution A: Weigh 2.05g of glucose, 0.055g of citric acid, and 0.42g of sodium chloride, add distilled water to 100ml, dissolve and sterilize by autoclaving, place in an environment at 4°C, and store for later use.

[0017] Solution B: 1g of canine albumin was weighed and dissolved in 10ml of normal saline to prepare a 10% canine albumin solution.

[0018] According to the ratio of 1:1, take the above solution A and solution B respectively, mix them well and put them in the refrigerator at 4°C for later use.

Embodiment 3

[0019] Example 3: Pre-transport treatment of MSCs.

[0020] Take the canine mesenchymal stem cells cultured in Example 1, transfer to the P3-P5 generation, adhere to the canine mesenchymal stem cells to 80-90%, digest with trypsin, centrifuge at 1200rpm for 5min, and discard the supernatant Resuspend the cells with normal saline, centrifuge again at 1200rpm for 5min and discard the supernatant. Add the special room temperature preservation solution for canine mesenchymal stem cells prepared in Example 2, and adjust the cell density to 5×10 5～6 / ml, take 1ml of cell suspension into a 2ml EP tube and seal it.

[0021] The prepared canine mesenchymal stem cell suspension can be directly injected locally or intravenously into the dog body. The suspension needs to be stored and transported in a light-proof environment at 4-25°C, and it is effective within 48 hours after preparation.

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PUM

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Abstract

The invention provides a special normal-temperature preservation fluid for a dog mesenchymal stem cell, which is prepared from dog blood albumin, normal saline, glucose, citric acid, and sodium chloride. The invention further provides an application method of the special normal-temperature preservation fluid for the dog mesenchymal stem cell. The method includes steps of adding the a dog mesenchymal stem cell performed with transportation pre-treatment in the special normal-temperature preservation fluid for the dog mesenchymal stem cell; adjusting cell density to 5x105-6 / ml; taking 1 ml cellsuspending fluid in 2 ml EP pipe and sealing. The special normal-temperature preservation fluid for the dog mesenchymal stem cell is easy to prepare and low in preparation cost; the fluid has the effect of storing the dog mesenchymal stem cell under the constant temperature, and can effectively prevent the dog mesenchymal stem cell from death or clustering.

Description

technical field [0001] The invention relates to the technical field of bioengineering. Background technique [0002] Mesenchymal Stem Cells (MSCs) are adult stem cells developed from mesoderm and present in various tissues of the body, which have the ability of self-renewal and multidirectional differentiation. In recent years, with the rise of stem cell therapy, MSC has been applied to the treatment of diseases such as blood diseases, skin trauma, liver and kidney injury, arthritis, myocarditis and cancer, and has achieved satisfactory results. [0003] The transportation of MSC is an important factor affecting its clinical popularization and application, and it is also a difficult problem that people have always wanted to break. Conventional MSC transportation includes ultra-low temperature transportation and room temperature adherent transportation. These two methods not only have high transportation costs, but also have many follow-up steps, which are not conducive to l...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/021
Inventor 王丙云詹小舒罗冬章石耀林陈志胜陈胜锋计慧琴
Owner FOSHAN UNIVERSITY
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