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Application of induced extracellular vesicles in preparation of preparations for prolonging life of mammals or treating or preventing aging

A mammalian, inducible technology, applied in the field of biomedicine, which can solve the problems of low exosome yield, time-consuming, and non-existent problems.

Pending Publication Date: 2022-01-21
EV CELL BIOTECH GUANGZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still many problems in the current exosome-based cell vesicle therapy, mainly in the complex and time-consuming process of exosome extraction and purification, high requirements for equipment and reagents, and low yield of physiological exosomes etc. These defects limit the clinical transformation and application of exosome therapy
[0005] There is no relevant research on effective extracellular vesicles for the preparation of drugs that extend the lifespan of mammals or treat or prevent aging-related diseases

Method used

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  • Application of induced extracellular vesicles in preparation of preparations for prolonging life of mammals or treating or preventing aging
  • Application of induced extracellular vesicles in preparation of preparations for prolonging life of mammals or treating or preventing aging
  • Application of induced extracellular vesicles in preparation of preparations for prolonging life of mammals or treating or preventing aging

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Example 1 Isolation and culture of BMMSCs

[0089] Excess CO was used according to the guidelines of the Animal Ethics Committee 2 Sacrifice the mice, remove the tibia and femur under aseptic conditions, peel off the muscles and connective tissue attached to them, further separate the metaphysis, expose the bone marrow cavity, and extract the volume fraction of 10% with a 10mL sterile syringe. The bone marrow cavity was washed repeatedly with PBS of fetal bovine serum, filtered through a cell strainer with a pore size of 70 μm, and centrifuged at 500 g for 5 min. After removing the supernatant, the cell pellet at the bottom was collected, resuspended in PBS, and centrifuged again at 500 g for 5 min to collect the final cell pellet. Then, the cells were sorted by flow cytometry, and bone marrow mesenchymal stem cells (BMMSCs) were sorted out by using CD34- and CD90+ as sorting criteria. Finally, the cells were resuspended in Dex(-) medium and inoculated in a 10cm diamet...

Embodiment 2

[0097] Example 2 Preparation of IEV derived from BMMSC

[0098] The MSCs (MSCs derived from bone marrow) cultured to the second generation in Example 1 were continued to be cultured with the medium (Dex(+) culture fluid) in Example 1 until the cells were confluent 80%-90%, washed with PBS for 2 Add serum-free medium containing 500nM STS (the medium is the medium in Example 1 with 500nM STS added), incubate at 37°C for 16-24h, collect the cell supernatant, and centrifuge at 800g for 10 hours at 4°C Minutes, the supernatant was collected and centrifuged at 2000g at 4°C for 10 minutes, and the supernatant was collected again and centrifuged at 16000g at 4°C for 30 minutes, and the obtained precipitate was IEV. The pellet was resuspended in 500 μl of PBS, centrifuged again at 16,000 g for 30 minutes at 4° C., and the washed IEV was obtained.

[0099] Preparation routes such as figure 2 shown.

Embodiment 3

[0103] The analysis of embodiment 3 IEVs

[0104] (1) Quantification of IEVs and analysis of membrane proteins

[0105] Utilize flow cytometry to carry out quantitative analysis to the IEVs that embodiment 2 obtains, measurement time point is the 1st, the 4th, the 8th, the 16th and the 24th h, the result shows 10 6 Each MSCs can produce 0.76×10 after induction to 1h, 4h, 8h, 16h and 24h respectively 8 pcs, 1.29×10 8 pcs, 1.95×108 pcs, 2.48×10 8 pcs, 3.14×10 8 IEVs, it can be seen that after induction to 24h, a single MSC can produce 300 IEVs ( image 3 ).

[0106] In addition, flow cytometry found that the particle diameter distribution of IEVs was concentrated below 1 μm, accounting for 94.97% ( Figure 4A ).

[0107] The results of side scattered light (SSC) analysis also showed that the scattered light intensity of IEVs was concentrated in the range below 1 μm ( Figure 4B ).

[0108] Further, the scattered light intensity of IEVs was analyzed by standardized small...

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Abstract

The invention discloses application of induced extracellular vesicles in preparation of preparations for prolonging the life of mammals or treating or preventing aging. Various embodiments prove that the induced extracellular vesicles have an anti-aging effect, can prolong the life of mammals, and have an effect of relieving senile alopecia. In addition, the invention also provides application of the induced extracellular vesicles in preparation of anti-aging, and / or repairing, and / or regeneration preparations for skin and / or skin appendages.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to the application of inducible extracellular vesicles in the preparation of preparations for prolonging the lifespan of mammals or treating or preventing aging. Background technique [0002] With the advent of the global aging society, the health of the elderly and the prevention and treatment of senile diseases have become important public health issues facing the international community. my country is the most populous country in the world, and it is also the country with the largest elderly population. "Aging society" is a serious problem facing our country. In order to solve the troubles brought about by the aging society, anti-aging medicine began to rise. Explore mechanisms of aging. Finding new targets and new treatments for anti-aging has become a major issue in life sciences. [0003] Aging is the lifelong deterioration of physiological integrity caused by externa...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K35/545A61P3/04A61P17/14A61P19/10A61P39/00A61P43/00C12N5/074C12N5/0775C12N5/0789
CPCA61K35/28A61K35/545A61P17/14A61P3/04A61P19/10A61P39/00A61P43/00C12N5/0663C12N5/0696C12N5/0647C12N2500/90C12N2501/999
Inventor 寇晓星施松涛
Owner EV CELL BIOTECH GUANGZHOU CO LTD
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