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Anti-aging modification method of adipose-derived mesenchymal stem cells and application of anti-aging modification method

A stem cell, fat-derived technology, applied in the field of biotechnology, can solve problems such as high aging burden, and achieve the effects of good treatment effect, high amplification efficiency, and improvement of treatment potential

Pending Publication Date: 2022-04-05
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, MSCs obtained from the elderly and patients with metabolic diseases will also have a higher burden of aging

Method used

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  • Anti-aging modification method of adipose-derived mesenchymal stem cells and application of anti-aging modification method
  • Anti-aging modification method of adipose-derived mesenchymal stem cells and application of anti-aging modification method
  • Anti-aging modification method of adipose-derived mesenchymal stem cells and application of anti-aging modification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 TRAF3 KD Modifications reduce TRAF3 protein expression levels in AMSC cells:

[0044] Detection of native AMSC, AMSC-Ctrl and AMSC-TRAF3 by Western Blot KD Protein expression level of TRAF3 in cells. And PCR method was used to detect the mRNA expression level of TRAF3 in the above cells.

[0045] Such as figure 1 shown by TRAF3 KD Modified mAMSC (mAMSC-TRAF3 KD ), the expression level of TRAF3 protein was significantly lower than that of control group mAMSC-Ctrl and natural mAMSC; similarly, TRAF3 KD Modified hAMSC (hAMSC-TRAF3 KD ), the TRAF3 protein expression level was also significantly lower than that of the control group hAMSC-Ctrl and natural hAMSC; while the TRAF3 mRNA level in AMSC cells derived from adult or mouse adipose tissue was significantly lower than that of the control group hAMSC-Ctrl and natural hAMSC; KD After modification, the differences were not significant.

Embodiment 2

[0046] Example 2 TRAF3 KD Modifications against AMSC cell senescence induced by culture passage:

[0047] Native AMSC, AMSC-Ctrl, and AMSC-TRAF3 from mouse and adult adipose tissue KD The cells were passed to the 9th passage (mAMSC) and the 25th passage (hAMSC) respectively, and the cell replication senescence model induced by culture passage was established.

[0048] Comparison of native AMSC, AMSC-Ctrl and AMSC-TRAF3 by SA-β-gal staining KD Cell staining. The staining of 5 visual fields was counted by light microscope, and the statistical analysis was carried out. The results showed that the positive rate of SA-β-gal blue staining of natural hAMSC was 32.5±3.4%, the positive rate of hAMSC-Ctrl blue staining was 37.5±4.4%, and hAMSC-TRAF3 KD The blue-stained positive rate of the cells was only 7.5±1.3%. The positive rate of SA-β-gal blue staining of natural mAMSC was 67.5±8.1%, the positive rate of blue staining of mAMSC-Ctrl was 71.5±9.7%, and the positive rate of mAMSC...

Embodiment 3

[0053] Example 3 TRAF3 KD Modifications inhibit chemotherapeutic drugs and radiation-induced AMSC senescence:

[0054] Cell senescence was induced by doxorubicin (Dox). Spread AMSC on a 6-well plate, culture overnight, then add 200nM Dox to treat for 48 hours, then change the normal medium and culture for another 6 days, collect cells, and use Western Blot method to detect natural AMSC, AMSC-Ctrl and AMSC-TRAF3 KD Expression levels of p21 and p16 proteins in cells.

[0055] The results showed that TRAF3 KD The modification can significantly reduce the increase of p21 and p16 protein expression in AMSC cells induced by chemotherapeutic drugs. In addition, in 5-FU, D-galactose, H 2 o 2 A similar phenomenon is also found in the induced cell senescence model and in the 137Csγ-ray-induced irradiated cell senescence model. showed that TRAF3 KD The modification was also protective against chemotherapeutic drug-induced AMSC senescence.

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Abstract

The invention provides an anti-aging modification method of adipose-derived mesenchymal stem cells (AMSC) and application of the anti-aging modification method, the anti-aging modification method is a modification method for intervening the aging of the adipose-derived mesenchymal stem cells (AMSC), the aging of the AMSC is inhibited through a modification strategy of epigenetically regulating the post-transcriptional level of a TRAF3 gene, replicative aging can be resisted based on the modification, and the anti-aging effect of the adipose-derived mesenchymal stem cells is improved. The cell senescence induced by chemotherapeutic drugs, toxins, oxidative stress, irradiation and the like, and the senescence load of cells derived from old individuals or metabolic disease individuals. Furthermore, the effect of TRAF3KD modification on improvement of the curative effect on the aging AMSC liver disease is determined through in-vivo experiments such as a mouse hepatic failure model and the like. The biological preparation for treating the hepatopathy, provided by the invention, is shorter in preparation period, and can eliminate adverse effects caused by AMSC aging related cell function damage, so that the application potential of the cell preparation in clinical treatment of the hepatopathy is improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an anti-aging modification method and application of adipose-derived mesenchymal stem cells, in particular to inhibiting the aging of adipose-derived mesenchymal stem cells by inhibiting translation of TRAF3 through epigenetic modification, thereby improving their proliferation ability and efficacy methods, and provide TRAF3 KD Modified AMSC (AMSC-TRAF3 KD ) in the preparation of biological agents for the treatment of liver diseases. . Background technique [0002] Liver failure is a type of severe damage to liver function caused by massive necrosis of liver cells caused by various etiologies, manifested as clinical syndromes such as hyperbilirubinemia, coagulation dysfunction, ascites, and hepatic encephalopathy. In my country, hepatitis B virus infection is the main factor leading to liver failure, and the acute attack of hepatitis B or the severe outcome of chronic hepatitis B ma...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N5/0775C12N15/867C12N15/12G01N33/68C12P19/02
Inventor 楼国华刘艳宁
Owner ZHEJIANG UNIV
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