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Preparation method of exosome, exosome prepared by preparation method and application of exosome

An exosome and autologous technology, applied in the preparation of exosomes, in the field of exosomes, to achieve the effect of promoting wound healing, reducing melanosis, and reducing traces

Pending Publication Date: 2022-01-11
三代康年(上海)医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the safety of adipose-derived mesenchymal stem cells is relatively high, stem cells with a complete structure still have a certain risk of tumorigenesis, and exosomes derived from adipose-derived mesenchymal stem cells take into account both safety and effectiveness

Method used

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  • Preparation method of exosome, exosome prepared by preparation method and application of exosome
  • Preparation method of exosome, exosome prepared by preparation method and application of exosome
  • Preparation method of exosome, exosome prepared by preparation method and application of exosome

Examples

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Effect test

Embodiment 1

[0065] This embodiment provides an exosome gel, which is prepared by the following preparation method:

[0066] (1) Take 20.01 mL of autologous abdominal adipose tissue (sample number S01025) and digest it with 20 mL of trypsin digestion solution, digest it at 37°C for 10 minutes, gently shake the digested cell suspension, and transfer it to a centrifuge tube; Digest again with 15mL of trypsin digestion solution, digest at 37°C for 5min; shake the digested suspension gently, transfer to a centrifuge tube, and collect the digested cell suspension;

[0067] Wherein, the trypsin digestion liquid includes: 0.1% trypsin, 0.1 mM EDTA solution with a pH of 8.0 as a solvent;

[0068] Centrifuge the collected digested cell suspension at 1000rpm for 3min, discard the supernatant; add 10mL of PBS buffer, shake and wash, then centrifuge at 1000rpm for 3min, discard the supernatant; repeat the washing step once to obtain adipocytes;

[0069] (2) The digested and cleaned adipocytes were cu...

Embodiment 2

[0075] This embodiment provides an exosome gel, which is prepared by the following preparation method:

[0076] (1) Take 21.43 mL of autologous abdominal adipose tissue (sample number S01026) and digest it with 20 mL of trypsin digestion solution, digest it at 37°C for 10 minutes, gently shake the digested cell suspension, and transfer it to a centrifuge tube; Digest again with 15mL of trypsin digestion solution, digest at 37°C for 5min; shake the digested suspension gently, transfer to a centrifuge tube, and collect the digested cell suspension;

[0077] Wherein, the trypsin digestion liquid includes: 0.1% trypsin, 0.1 mM EDTA solution with a pH of 8.0 as a solvent;

[0078] Centrifuge the collected digested cell suspension at 1000rpm for 3min, discard the supernatant; add 10mL of PBS buffer, shake and wash, then centrifuge at 1000rpm for 3min, discard the supernatant; repeat the washing step once to obtain adipocytes;

[0079] (2) The digested and cleaned adipocytes were cu...

Embodiment 3

[0085]This embodiment provides an exosome gel, which is prepared by the following preparation method:

[0086] (1) Take 16.23 mL of autologous abdominal adipose tissue (sample number S01028) and digest it with 15 mL of trypsin digestion solution, digest it at 37°C for 10 minutes, gently shake the digested cell suspension, and transfer it to a centrifuge tube; Digest again with 15mL of trypsin digestion solution, digest at 37°C for 5min; shake the digested suspension gently, transfer to a centrifuge tube, and collect the digested cell suspension;

[0087] Wherein, the trypsin digestion liquid includes: 0.1% trypsin, 0.1 mM EDTA solution with a pH of 8.0 as a solvent;

[0088] Centrifuge the collected digested cell suspension at 1000rpm for 3min, discard the supernatant; add 10mL of PBS buffer, shake and wash, then centrifuge at 1000rpm for 3min, discard the supernatant; repeat the washing step once to obtain adipocytes;

[0089] (2) The digested and cleaned adipocytes were cul...

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Abstract

The invention provides a preparation method of an exosome, the exosome prepared by the preparation method and application of the exosome. The preparation method comprises the following steps: (1) digesting autologous adipose tissues with trypsin digestive juice, and carrying out cleaning to obtain adipose cells; (2) culturing the adipose cells obtained in the step (1) in a culture medium to obtain a cell suspension; and (3) mixing the cell suspension obtained in the step (2) with tranexamic acid, then, carrying out culturing, and carrying out collecting to obtain the tranexamic-acid-modified exosome generated by autologous mesenchymal adipose-derived stem cells. Effective components of the exosome preparation comprise the exosome generated by the autologous mesenchymal adipose-derived stem cells and modification of the tranexamic acid, and the exosome preparation for repairing burn wounds needs to be capable of promoting wound healing, reducing melanin deposition of the wounds and reducing traces after wound healing.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing exosomes, exosomes prepared therefrom and applications thereof. Background technique [0002] Skin burns are open pathological lesions, and burns can cause damage to skin structure and function. Skin healing after burns is a complex physiological process. A variety of tissue cells are required to participate in the process, such as epithelial cells, endothelial cells, immune cells, fibroblasts, etc. Burn wounds are exposed to a complex microbial environment due to the presence of a large amount of burn toxins. Appropriate participation of inflammatory cells can reduce the risk of infection, but excessive inflammatory responses will affect the healing of burn wounds. [0003] For patients with extensive burns, due to the lack of autologous skin sources, autologous skin transplantation is limited; the skin from allogeneic sources is not only limited,...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775A61K35/28A61K9/06A61K47/36A61P17/02
CPCC12N5/0653C12N5/0667A61K35/28A61K9/06A61K47/36A61P17/02C12N2509/00
Inventor 李海源卢俊锋陈晓冬
Owner 三代康年(上海)医疗科技有限公司
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