Application and preparation method of autologous fibroblast-sourced freeze-dried exosome powder

A technology of fibroblasts and exosomes, applied in the field of medical biology, can solve the problems of short-lived, difficult storage and transportation of stem cells, and low differentiation rate, and achieve the effects of promoting synthesis, anti-wrinkle and anti-oxidation effects, and high safety

Inactive Publication Date: 2018-12-18
浙江卫未生物医药科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, at present, in the repair of skin wounds, stem cells mainly rely on paracrine to promote the repair of skin wounds, and the transplantation of stem cells to the injured site has a low differentiation rate and a short-term effect.
In addition, the use of stem cells has limitations such as difficulties in storage and transportation, ethics, tumorigenic mutations, and immune rejection. Therefore, it is very important to find a method that has the curative effect of stem cells and can overcome the shortcomings of stem cell therapy.

Method used

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  • Application and preparation method of autologous fibroblast-sourced freeze-dried exosome powder
  • Application and preparation method of autologous fibroblast-sourced freeze-dried exosome powder
  • Application and preparation method of autologous fibroblast-sourced freeze-dried exosome powder

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preparation example Construction

[0025] A preparation method of exosome freeze-dried powder derived from autologous fibroblasts, comprising the following steps:

[0026] (1) Obtaining skin-derived fibroblasts:

[0027] Take 0.02g of the skin behind the ears of a healthy person (age 25-35 years old), digest it with 0.25% dispase for 1.5h, remove the tissue, fur and fat layer and leave the dermis, then digest it with 0.25% type 1 collagenase for 1.5h, 300× Centrifuge at g10min, resuspend the pellet with 4ml of fibroblast serum-free complete medium, add to a T-25 cell culture flask, place at 37°C, 5% CO 2 Cultivate in an incubator, change the medium with half of the fibroblast serum-free basal medium every 3 days, cell colonies appear after 4 days of culture, the cells are overgrown in 12-15 days, and pass passage, the cells are in the shape of a long spindle (such as figure 1 shown), is the morphology of fibroblasts.

[0028] (2) Cell expansion and cell identification: When the cells in the step (1) cover mor...

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Abstract

The invention relates to application and a preparation method of an autologous fibroblast-sourced freeze-dried exosome powder. The autologous fibroblast-sourced freeze-dried exosome powder is appliedinto wrinkle resistance and oxidation resistance, and the application method is as follows: normal saline is used for dissolving the autologous fibroblast-sourced freeze-dried exosome powder into 0.5percent by weight of solvent, so that injection is obtained, and the injection is injected deep into deep stratum of the dermis by a microneedle. The preparation method of the freeze-dried exosome powder comprises four steps, i.e., obtainingment of skin-sourced fibroblasts, cell amplification and cell identification, separation and identification of exosome and freeze-drying of exosome. Since theinvention adopts the autologously sourced exosome to resist wrinkles and oxidation, the autologous fibroblast-sourced freeze-dried exosome powder is easy to store and transport, ethical problems do not exist, moreover, safety is high, and the wrinkle-resistant and oxidation-resistant effect is good.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to an application and preparation method of exosome freeze-dried powder derived from autologous fibroblasts. Background technique [0002] Wrinkles are due to the lack of collagen, hyaluronic acid, elastic fibers and other substances in the dermis of the skin, which cannot support the normal structure of the skin, and the epidermis of the skin collapses to form gullies. These important substances are all secreted by fibroblasts. That is to say, fibroblasts are the main effector cells in the process of wrinkle formation. Collagen makes up 70-80% of the dry weight of the skin and contributes to the stability and structural integrity of the tissue. A large number of studies have proved that the gradual decline of collagen synthesis in the dermis leads to the formation of wrinkles and aging skin, collagen directly affects skin tension, and the maintenance of collagen structure is relate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/99A61K8/98A61K8/02A61Q19/08
CPCA61K8/022A61K8/981A61K8/99A61K2800/782A61Q19/08
Inventor 张雪芬陈锦阳刘军权
Owner 浙江卫未生物医药科技有限公司
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