Activity detection method of stem cell telomere repair active substance with good stability

An active substance and activity detection technology, applied in the A61 medical field, can solve problems such as long detection time, difficult quantitative analysis, and limited detection sample range, and achieve the effects of improving stability, stable extraction effect, and improving speed and effect

Pending Publication Date: 2022-06-17
华夏源细胞工程集团股份有限公司
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AI Technical Summary

Problems solved by technology

[0003] Common methods for detecting the activity of telomere repair active substances in human mesenchymal stem cells mainly include TRAP method, TRAP-ELISA method and TRAP silver staining method, etc. However, the previous detection methods have the following problems, such as the limited ra...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 The first aspect provides a detection method for the activity of the activity of the activity of the reconciliation of the cobies of the cobies of the albumin telomere of the world. S2: Configure the reverse transcription reaction solution and use the PCR instrument to perform a reversal transcription reaction; S3: configure the QPCR reaction solution, and use the QPCR instrument to perform fluorescent quantitative reactions; S4: The relative activity judgment of telomere restoring activity.

[0046] The specific operation method of S1 steps in this embodiment includes the following steps: (1) Collect the germinated stem cells for measuring the metering of sodium chloride injection solution 3 × 10 6 This, 400g centrifugal 5 minutes, go to the upper clearing, and leave the accuracy of the germinal stem cells; The extraction aids (sulfur sulfur glycol and β-sulfur ethanol with a volume ratio of 4: 6) are added to 100 μl of cell cracking buffer, and then the vortex i...

Embodiment 2

[0058] The specific implementation method of this embodiment is the same as the implementation example 1. The difference is that the extract agent of the extract agent of the benzo -sulfonal sulfonyl fluorine cell cracking buffer is 0.4 μl, and the extraction auxiliary agent that is diluted by 20 times the cell crack is 0.7 μL.

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PUM

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Abstract

The invention discloses an activity detection method for stem cell telomere repair active substances with good stability, which is characterized by comprising the following steps: S1, preparing a cell lysis solution, and performing lysis on mesenchymal stem cells to extract target protein; s2, preparing a reverse transcription reaction solution, and carrying out a reverse transcription reaction by adopting a PCR instrument; s3, preparing a QPCR reaction solution, and carrying out a fluorescent quantitative reaction by adopting a QPCR instrument; and S4, judging the relative activity of the telomere repairing active substance. The phenylmethylsulfonyl fluoride, the beta-mercaptoethanol and the dithiothreitol are compounded and added into the cell lysis solution, and the volume ratio of the phenylmethylsulfonyl fluoride, the beta-mercaptoethanol and the dithiothreitol in the cell lysis solution is limited, so that the cell lysis and reaction efficiency and quality are effectively improved; the time required for detecting the activity of the mesenchymal stem cell telomere repair active substance is fully shortened, the stability of the whole detection method is obviously improved, and the accuracy is higher.

Description

Technical field [0001] The invention involves the A61 medical field in the IPC classification, especially the active detection method of a good stable stem cell telomere restoration. Background technique [0002] Medium -sized stem cells are currently attracting attention of stem cells with high self -renewal and multi -directional differentiation potential. In the related research of mesenchymal stem cells, the research on the active activity of telomeres of mesenchymal stem cells occupy an important research. Research status. Because telomrase is a kind of enzyme that is responsible for telomey in the mesenchymal stem cells, it controls the number of stem cell differentiation and division, and its activity has a clear decisive effect on the proliferation of stem cells. [0003] The method of common human essential stem cell telomere restoration of the activity detection method mainly includes the Trap method, the Trap-Elisa method, and the trap silver dyeing method. However The...

Claims

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Application Information

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IPC IPC(8): C12Q1/48C12Q1/6806C12Q1/6851
CPCC12Q1/485C12Q1/6806C12Q1/6851G01N2333/9128C12Q2527/125C12Q2523/10C12Q2521/113C12Q2531/113C12Q2563/107
Inventor 朱灏
Owner 华夏源细胞工程集团股份有限公司
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