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Method for quickly reaerating cells under anaerobic environment

A cell-based and rapid technology, applied in the field of biomedicine, can solve the problems of complex operation, high cost, and long experiment time, and achieve the effect of high biological safety, strong operability, and simple method

Inactive Publication Date: 2017-08-15
SHANGHAI NAT ENG RES CENT FORNANOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The purely physical reoxygenation model has a wide range of applications and flexible design, but its specificity is not high, the experimental results are greatly affected by the experimental conditions, and the operation is complicated, the cost is high, and the experimental time is long

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Dissolve 10 parts of fluorocarbons in 100 parts of ethanol, add 1 part of heptane and cyclobutyl ether at the same time, stir and mix thoroughly for 10 minutes, and prepare an ethanol solution of oxygen carrier.

[0023] 2. Take normal adherent cells and place them in a three-gas incubator with 1% oxygen concentration. After culturing for 24 hours, suck out the original medium, replace it with a medium containing oxygen carrier, add Triton X-100, and in normal oxygen concentration for 2 hours.

[0024] 3. Aspirate the excess oxygen carrier medium, add 100μ, 10% CCK-8 reagent, incubate in a constant temperature incubator at 37°C for 2 hours, detect its absorbance with a microplate reader, and measure the activity of the cells.

[0025] Test result: OD value=0.43.

Embodiment 2

[0027] 1. Dissolve 10 parts of fluorocarbons in 100 parts of ethanol, add 10 parts of cyclopentane and cyclohexyl ether at the same time, stir and mix thoroughly for 10 minutes, and prepare an ethanol solution of oxygen carrier.

[0028] 2. Take normal adherent cells and place them in a three-gas incubator with 1% oxygen concentration. After culturing for 48 hours, suck out the original medium, replace it with a medium containing oxygen carrier, add Tu-80, and in normal oxygen concentration Incubate for 12 hours.

[0029] 3. Aspirate the excess oxygen carrier medium, add 100μ, 10% CCK-8 reagent, incubate in a constant temperature incubator at 37°C for 2 hours, detect its absorbance with a microplate reader, and measure the activity of the cells.

[0030] Test result: OD value=0.67.

Embodiment 3

[0032] 1. Dissolve 10 parts of fluorocarbons in 100 parts of ethanol, add 10 parts of cyclopentane and cyclohexyl ether at the same time, stir and mix thoroughly for 10 minutes, and prepare an ethanol solution of oxygen carrier.

[0033] 2. Take normal adherent cells and place them in a three-gas incubator with 10% oxygen concentration. After culturing for 24 hours, suck out the original medium, replace it with a medium containing oxygen carriers, add Tu-80, and in normal oxygen concentration Incubate for 24 hours.

[0034] 3. Aspirate the excess oxygen carrier medium, add 100μ, 10% CCK-8 reagent, incubate in a constant temperature incubator at 37°C for 2 hours, detect its absorbance with a microplate reader, and measure the activity of the cells.

[0035] Test result: OD value=0.52.

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Abstract

The invention relates to a method for quickly reaerating cells under an anaerobic environment. The method comprises the following steps: preparing an oxygen carrier, culturing cells and detecting cells: sucking redundant oxygen carrier culture medium, adding 100 microliter of 10% 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-benezene dithionate)-2H-tetrazole monosodium (CCK-8) reagent, incubating for 2-4 hours in a constant temperature incubation box at 37 DEG C, using a microplate reader for detecting light absorbance thereof and determining the cell activity. The oxygen carrier can enrich a large amount of oxygen, is used as an oxygen supplier and can supply oxygen for the cells under the condition of insufficient air oxygen. According to the invention, no external matter is added, the biological safety is high, the toxicity is low, the method is simple and the operability is strong.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for rapid reoxygenation of cells in a hypoxic environment, and a method for rapid reoxygenation of cells in a hypoxic environment by using fluorocarbons. Background technique [0002] Ischemic diseases seriously endanger human health. Therefore, shortening the tissue ischemic time and restoring blood flow as soon as possible are the most effective measures to prevent and treat ischemic injury. There are many hypoxia-reoxygenation models reported so far, including purely physical models and purely chemical models. Purely physical reoxygenation models are more common, and commonly used methods include mixed gas culture method, simulated hypoxia / reperfusion fluid culture method, nitrogen saturated culture method, liquid paraffin covering method, etc. The purely physical reoxygenation model has a wide range of applications and flexible design, but its specificity is not high, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
Inventor 何丹农朱君王杰易帆乔宇金彩虹
Owner SHANGHAI NAT ENG RES CENT FORNANOTECH
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