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Preparing method and application of human mesenchymal stem cell source exosome bio-active preparation

A technology of stem cells and exosomes, applied in the field of biomedicine, can solve the problem of insignificant anti-aging effect, achieve large market prospects and economic value, and save time and cost.

Inactive Publication Date: 2017-07-04
深圳市赛欧细胞技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For this reason, the technical problem to be solved by the present invention is to overcome the technical bottleneck that the existing cosmetic preparations have no obvious anti-aging effect and rarely use exosomes as raw materials, so as to propose a slowing down method of human mesenchymal stem cell-derived exosomes. The preparation method of release cosmetic preparation

Method used

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  • Preparing method and application of human mesenchymal stem cell source exosome bio-active preparation
  • Preparing method and application of human mesenchymal stem cell source exosome bio-active preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 This example discloses a preparation method of cosmetic preparations, the specific steps are as follows:

[0038] 1) Weigh 200 mg of chitosan and dissolve it in 100 mL of 0.5% dilute acetic acid to obtain a chitosan solution.

[0039] 2) Weigh 5 mg of human mesenchymal stem cell-derived exosome freeze-dried powder, 10 mg of glucosaminoglycan, and 10 mg of hyaluronic acid, and dissolve them in 50 mL of glycerol to obtain an exosome solution.

[0040] 3) Slowly add the exosome solution into the chitosan solution dropwise under continuous stirring, and react for 30 minutes to obtain the chitosan nanoparticle suspension of the exosome.

[0041] 4) The suspension was centrifuged at 4 times at high speed (10000r / min) for 30 minutes, the precipitate was collected, washed 3 times with water for injection, and dried to obtain the human mesenchymal stem cell-derived exosome sustained-release cosmetic preparation.

Embodiment 2

[0042] Example 2 This example discloses the method for preparing the freeze-dried powder of exosomes described in Example 1

[0043] Take exosomes, add mannitol (1%-15%), mix well, filter (0.22 μm), sub-package, and then place in an ultra-low temperature refrigerator (-80°C) for 10h. Then, freeze-dry (-50° C., 24 h) under a vacuum of 10 Pa to obtain a freeze-dried powder of human mesenchymal stem cell-derived exosomes.

Embodiment 3

[0044] Example 3 This example discloses the preparation method of exosomes described in Example 2

[0045] A. Human MSC culture and pretreatment process:

[0046] 1. Materials and reagents

[0047] 1) For MSCs from human umbilical cord, placenta, bone marrow, fat or other sources, the number of passages can be within 6 generations.

[0048] 2) Human MSC serum-free medium

[0049] 3) alpha-MEM

[0050] 4) Recombinant human interferon

[0051] 5) Recombinant human EPO

[0052] 6) Recombinant human PDGF-BB

[0053] 7) Petri dish (diameter 150mm)

[0054] 8) Other consumables

[0055] B. Cell culture process

[0056] 1) Digest and count human subcultured MSCs, suspend them in MSC serum-free complete medium, and adjust the cell concentration to 1.0×10 6 / ml.

[0057] 2) according to 10 6 Inoculate in a 150mm large Petri dish and culture until the cell confluency reaches 70-80%. It usually takes 2-3 days.

[0058] 3) Aspirate the liquid in the petri dish, and wash the p...

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Abstract

The invention belongs to the field of biological medicine, and particularly relates to a preparing method and application of a human mesenchymal stem cell source exosome bio-active preparation. The method comprises the steps of weighing and dissolving 50-400 mg of chitosan in 25-200 mL of dilute acetic acid to obtain chitosan solution, and weighing and dissolving 1-10 mg of human mesenchymal stem cell source exosome freeze-dried powder, 2-20 mg of glucosaminoglycans and 2-20 mg of hyaluronic acid in 10-100 mL of glycerin to obtain exosome solution; adding the exosome solution to the chitosan solution dropwise to obtain chitosan nanoparticle suspension of exosome; and conducting centrifugal treatment on the suspension, collecting sediment, and conducting washing and drying treatment to obtain the beautifying preparation. The problem of skin aging is solved, frequent use is avoided, adverse reaction of beautifying preparations is reduced, the effective concentration of active ingredients can be maintained for a long time, and the best treatment effect can be realized with the minimum dose.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a preparation method and application of a human mesenchymal stem cell-derived exosome bioactive preparation. Background technique [0002] It is well known that normal cells, as well as tumor cells, require cell-to-cell communication via cell-secreted vectors to maintain physiological state and activity. Among them, exosomes are an important intercellular communication carrier that can be produced by all types of cells and have been found in various body fluids, including blood, lymph, urine and milk. The results of nearly three decades of research have shown that exosomes carry abundant cell-derived substances, including liposomes, proteins, mRNA, non-coding RNA, and even DNA fragments. Exosomes are not simple cellular molecular garbage, they can be taken up by other cells. Therefore, exosomes can transfer biological information between cells, thus playing an important r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P17/00A61P17/18C12N5/0775
CPCA61K35/28C12N5/0663C12N5/0665C12N5/0667C12N5/0668C12N2501/135C12N2501/14C12N2501/24
Inventor 刘家权张涤平郭子宽陈贤光钱开诚鲁立
Owner 深圳市赛欧细胞技术有限公司
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