Experimental method for rapidly detecting activity of human mesenchymal stem cell IDO1

A mesenchymal stem cell and IDO1 technology, applied in the direction of cell culture active agent, animal cells, vertebrate cells, etc., can solve the problems of human mesenchymal stem cell immune regulation function evaluation error, cell adhesion, affecting test results, etc., to improve the evaluation Accuracy, improved accuracy, high stability effect

Pending Publication Date: 2021-08-20
华夏源(上海)生命科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, while detecting the activity of indoleamine 2,3-dioxygenase 1, it is necessary to cultivate human mesenchymal stem cells. At this stage, in the process of culturing human mesenchymal stem cells, there is a problem of cell adhesion due to high cell seeding density. Serious and other phenomena will affect the test results and lead to errors in the evaluation of the immune regulation function of human mesenchymal stem cells

Method used

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  • Experimental method for rapidly detecting activity of human mesenchymal stem cell IDO1
  • Experimental method for rapidly detecting activity of human mesenchymal stem cell IDO1
  • Experimental method for rapidly detecting activity of human mesenchymal stem cell IDO1

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Embodiment 1

[0081] An experimental method for rapidly detecting IDO1 activity of human mesenchymal stem cells, comprising the following steps:

[0082] S1: Human mesenchymal stem cell culture:

[0083] 1) Human mesenchymal stem cells were cultured in MM3 medium for 24 hours;

[0084] 2) Then half of the culture medium was replaced with MM3 medium containing interferon-γ for culture, and the other half was cultured with MM3 medium for 24 hours;

[0085] S2: Take the supernatant to detect the activity of indoleamine 2,3-dioxygenase 1:

[0086] Take the cultured supernatant, centrifuge, take 160 μL supernatant to react with 40 μL mass concentration of 30% trichloroacetic acid solution, centrifuge after the reaction, take the supernatant and mass concentration of 2% p-dimethylaminobenzaldehyde solution and mix in equal volumes, statically The absorbance value at 492nm was detected after setting. At the same time, use kynurenine standard solution, mix it with 2% p-dimethylaminobenzaldehyde ...

Embodiment 2

[0098] An experimental method for rapidly detecting IDO1 activity of human mesenchymal stem cells, comprising the following steps:

[0099] S1: Human mesenchymal stem cell culture:

[0100] 1) Human mesenchymal stem cells were cultured in MM3 medium for 24 hours;

[0101] 2) Then replace the MM3 culture medium and continue for 24 hours;

[0102] S2: Take the supernatant to detect the activity of indoleamine 2,3-dioxygenase 1:

[0103] Take the cultured supernatant, centrifuge, take 160 μL supernatant to react with 40 μL mass concentration of 30% trichloroacetic acid solution, centrifuge after the reaction, take the supernatant and mass concentration of 2% p-dimethylaminobenzaldehyde solution and mix in equal volumes, statically The absorbance value at 492nm was detected after setting. At the same time, use kynurenine standard solution, mix it with 2% p-dimethylaminobenzaldehyde solution in equal volume, and measure the absorbance value at 492nm after standing still.

[010...

Embodiment 3

[0114] An experimental method for rapidly detecting IDO1 activity of human mesenchymal stem cells, comprising the following steps:

[0115] S1: Human mesenchymal stem cell culture:

[0116] 1) Human mesenchymal stem cells were cultured in MM3 medium for 24 hours;

[0117] 2) Then the culture medium was replaced with MM3 medium containing interferon-γ for 24 hours;

[0118] S2: Take the supernatant to detect the activity of indoleamine 2,3-dioxygenase 1:

[0119] Take the cultured supernatant, centrifuge, take 160 μL supernatant to react with 40 μL mass concentration of 30% trichloroacetic acid solution, centrifuge after the reaction, take the supernatant and mass concentration of 2% p-dimethylaminobenzaldehyde solution and mix in equal volumes, statically The absorbance value at 492nm was detected after setting. At the same time, use kynurenine standard solution, mix it with 2% p-dimethylaminobenzaldehyde solution in equal volume, and measure the absorbance value at 492nm a...

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Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to an experimental method for rapidly detecting the activity of a human mesenchymal stem cell IDO1. The experimental method for rapidly detecting the activity of the human mesenchymal stem cell IDO1 comprises the following steps of: S1, culturing human mesenchymal stem cells; and S2, taking supernatant, and detecting the activity of indoleamine 2, 3-dioxygenase 1. By improving and optimizing a human mesenchymal stem cell culture method, the phenomena of cell adherence, quantity reduction and the like caused by high-density inoculation before cells are solved, so that the accuracy of a detection result is improved, and the accurate estimation and evaluation of the immune regulation and control function of the human mesenchymal stem cells are improved; and by determining a kynurenine solution, a trichloroacetic acid solution and a dimethylaminobenzaldehyde solution and selecting the mass concentration of the solutions, the evaluation accuracy of the function of the human mesenchymal stem cells is further improved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and more specifically relates to an experimental method for rapidly detecting IDO1 activity of human mesenchymal stem cells. Background technique [0002] Human mesenchymal stem cells are a kind of pluripotent stem cells, which have all the common characteristics of stem cells - self-renewal and multi-directional differentiation capabilities. They are also most widely used in clinical practice. Combined application with hematopoietic stem cells can improve the success rate of transplantation and accelerate hematopoietic reconstruction . In current applications, the immune regulation function of human mesenchymal stem cells has been regarded as an important key role. When human mesenchymal stem cells play an immune role, they also interact with various immune cells or secrete different active molecules to play their immune role. At present, it is considered that indoleamine 2,3-dioxygenase 1...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/26C12N5/0775
CPCC12N5/0662C12N2501/24C12Q1/26
Inventor 朱灏
Owner 华夏源(上海)生命科技有限公司
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