Skin epidermal cell oxidation resistance of mesenchymal stem cell exosome and preparation method of mesenchymal stem cell exosome preparation

A technology of stem cells and epidermal cells, which is applied in skin care preparations, skin diseases, cosmetic preparations, etc., can solve the problems of secondary skin damage, difficulty in polymerization, and limited anti-oxidative stress effects, and achieves growth promotion and improvement. Enzyme activity, protective effect against oxidative damage

Inactive Publication Date: 2019-03-15
陕西九州生物医药科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] (1) At present, there is no objective experimental evidence for the anti-oxidation or aging of exosomes from human umbilical cord mesenchymal stem cells;
[0006] (2) Individual inventions mention that exosomes from human umbilical cord mesenchymal stem cells can improve the activity of SOD enzymes in cells, but the anti-oxidative stress effect is too limited;
[0007] (3) Stem cell exosomes are used as biological agents, and direct skin injection may cause secondary damage to the skin;
[0008] (4) The main components of exosomes are small molecular substances, which are difficult to aggregate when applied directly, and have limited anti-oxidation or aging effects on the skin

Method used

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  • Skin epidermal cell oxidation resistance of mesenchymal stem cell exosome and preparation method of mesenchymal stem cell exosome preparation
  • Skin epidermal cell oxidation resistance of mesenchymal stem cell exosome and preparation method of mesenchymal stem cell exosome preparation
  • Skin epidermal cell oxidation resistance of mesenchymal stem cell exosome and preparation method of mesenchymal stem cell exosome preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Collection of supernatant of human umbilical cord mesenchymal stem cells and extraction of exosomes

[0047] 1. Culture of human umbilical cord mesenchymal stem cells:

[0048] The umbilical cord tissue of full-term healthy infants was collected, placed in a sterile bottle filled with PBS (containing 100 U / mL penicillin and streptomycin), and processed within 4 hours. Peel off the veins and arteries of the umbilical cord, strip off Wharton's jelly, and cut into 2mm 3 Spread evenly in the culture flask, add a small amount of complete medium to keep it moist, the medium is DMEM / F12 plus 15% FBS, add enough medium after 24 hours, and change the medium every 3 days after that, the cells grow until they are confluent For subculture, the culture medium after passage uses DMEM / F12 plus 10% FBS.

[0049] 2. Extraction of exosomes:

[0050] Exosomes were extracted from the supernatant of human umbilical cord mesenchymal stem cells of the 3rd to 5th passages. After ...

Embodiment 2

[0056] Example 2. In vitro safety experiment of human umbilical cord mesenchymal stem cell exosomes

[0057] CCK-8 method was used to detect the effect of exosomes from human umbilical cord mesenchymal stem cells on the growth of epidermal cells.

[0058] Test sample: the human umbilical cord mesenchymal stem cell exosome suspension prepared in Example 1.

[0059] Test organism: Human epidermal cells.

[0060] Specific steps:

[0061] (1) Epidermal cells in the logarithmic growth phase were inoculated into 96-well plates, 5000 cells / well, and divided into control group and exosome groups with different concentrations (1 μg / mL, 5 μg / mL, 10 μg / mL, 30 μg / mL ,50μg / mL).

[0062] (2) 24 hours after inoculating the cells, different amounts of exosomes were added according to the groups. On the following 1d, 3d, 5d and 7d, the old medium in the wells of the culture plate was discarded, and 10% CCK-8 reagent was added. The medium was incubated at 37°C for 2 hours, and then the abso...

Embodiment 3

[0065] Example 3. In vivo safety experiment of human umbilical cord mesenchymal stem cell exosomes

[0066] 1. Test sample: the human umbilical cord mesenchymal stem cell exosome suspension prepared in Example 1.

[0067] 2. Test organism: male Kunming mice.

[0068] 3. Specific steps:

[0069] (1) 12 male Kunming mice weighing 35-45 g were selected, and the back hair was removed with a depilatory agent.

[0070] (2) Randomly divided into normal group, normal saline group and different doses of exosome groups (5-10μg / vessel / day, 20-50μg / vessel / day), 3 in each group, exosome group with daily back A certain amount of exosomes was injected subcutaneously, and 20 μL of normal saline was injected into the normal saline group.

[0071] (3) Continue to inject for 7 days, and observe the changes of the back skin every day.

[0072] (4) It can be seen from Figure 2 that the mice injected with exosomes were observed after 7 days, and compared with the normal group, there was no abno...

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Abstract

The invention discloses skin epidermal cell oxidation resistance of a mesenchymal stem cell exosome and a preparation method of a mesenchymal stem cell exosome preparation. The safety and effectiveness of skin epidermal cell oxidation resistance of a mesenchymal stem cell exosome are verified. Experimental results show that the mesenchymal stem cell exosome is capable of effectively promoting proliferation of epidermal cells, inhibiting oxidative damage of epidermal cell DNA and improving SOD (Superoxide Dismutase) enzymatic activity of the epidermal cells after oxidative stress, and is safe and non-toxic. Therefore, the invention provides a preparation method of a skin epidermal cell oxidation resistant preparation of the mesenchymal stem cell exosome.

Description

technical field [0001] The present invention relates to a new application of stem cell exosomes, in particular to the anti-oxidation properties of mesenchymal stem cell exosomes and the preparation method of preparations thereof, especially the preparation method of human umbilical cord mesenchymal stem cell exosomes new use. It belongs to the field of stem cells and transformation. Background technique [0002] Extracellular vesicles (EVs) are considered efficient vehicles for intercellular communication due to their ability to deliver proteins, lipids, and nucleic acids, thereby affecting various physiological and pathological functions of recipient and parent cells. Exosomes in EVs Because it includes a large number of proteins, lipids, mRNA, miRNA, non-coding RNA and major histocompatibility complexes, it plays a key regulatory role in tissue repair, immune response and oxidative stress. The skin can produce oxidative stress under ultraviolet radiation or toxic subst...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61P17/18A61K8/99A61K8/98A61Q19/00A61Q19/08
CPCA61K8/982A61K8/99A61K9/0014A61K35/28A61K2800/78A61Q19/00A61Q19/08A61P17/18
Inventor 龙乾发戴博田丽军聂苏秦何婷婷鲜盼盼李敏王甜张博
Owner 陕西九州生物医药科技集团有限公司
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