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

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...

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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 range of detection samples (not applicable to fresh detection of mesenchymal stem cells recovered after repeated cryopreservation), the detection time is relatively long and the number of detection samples is limited, the experimental results are unstable and difficult to quantitatively analyze, etc.