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Application of combination of polypeptide conjugates and epidermal stem cell exosomes in medicine and cosmetics

An epidermal stem cell and polypeptide coupling technology, applied in the field of biomedicine, can solve the problem of insufficient research and achieve the effect of inhibiting melanin synthesis

Active Publication Date: 2020-05-08
广州市新纪元化妆品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] However, judging from the disclosed skin cosmetics for exosomes, on the one hand, the research is not very sufficient, and on the other hand, there is still a lot of room for improvement in cosmetic effects

Method used

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  • Application of combination of polypeptide conjugates and epidermal stem cell exosomes in medicine and cosmetics
  • Application of combination of polypeptide conjugates and epidermal stem cell exosomes in medicine and cosmetics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Culture of Mouse Epidermal Stem Cells

[0033]Inoculate mouse epidermal stem cells (mouse epidermal stem cells, product number BJ-ATCC0672, Shanghai Bangjing Industrial Co., Ltd.) on a 25mm culture flask growing NIH3T3 cells, add special medium: DMEM:HAM F-12=3:1 Mixed with 10% FBS, 20ng / ml EGF, 5ug / ml transferrin, 1.8×10 -4 mo1 / L adenine, 5ug / ml insulin and 0.4ug / ml hydrocortisone, pH = 7.0-7.4 Change the medium every 2 days, keep the culture environment at humidity >95%, temperature 37°C, C0 2 Concentration 5%. After the cells are 80% confluent, they can be passaged. Firstly, NIH3T3 cells are removed with 0.02% EDTA, and then digested with 0.25% trypsin and 0.02% EDTA for passage. This culture and expansion process was repeated for 3 times to complete the culture of a large number of epidermal stem cells.

Embodiment 2

[0034] Example 2 Extraction of Epidermal Stem Cell Exosomes

[0035] The supernatant of the cells cultured in Example 1 was divided into 50 ml centrifuge tubes, centrifuged at 3000 rpm at room temperature for 20 min, and the cells and their cell debris were removed. Collect the supernatant after centrifugation, pass through a 0.22 μm filter membrane to remove large particles and vesicles, add PEG 6000 reagent with a final concentration of 0.5% trehalose and a final concentration of 8%, and let stand overnight at 4 °C. Centrifuge the overnight PEG 6000 cell supernatant mixture at 3000rpm for 30min. It can be seen that there are white flocculent precipitates on the bottom and wall of the centrifuge tube. Discard the supernatant of the centrifuge tube and place the centrifuge tube upside down on absorbent paper. Air dry for 5-10 minutes. According to the amount of precipitate, according to the volume ratio of 0.1g:10mL, add an appropriate amount of PBS or double distilled water ...

Embodiment 3

[0036] Example 3 The whitening effect of the combination of exosomes and polypeptide compounds derived from epidermal stem cells

[0037] The whitening effect of the combination of exosomes and compounds was determined by the degree of inhibition of melanogenesis in mouse melanoma. Cultured melanoma cells (A875 (human melanoma cells), product number: EY-X0770, Shanghai Yiyan Biotechnology Co., Ltd.) were mixed with 1×10 5 The cell / well density was seeded in a 96-well plate, and 10 mg / L of the compound of formula (I) was added to the exosomes with a final concentration of 30 and 50 ug / ml (This compound was developed by the inventor in the early stage, CN110680928A Chinese formula (II) is disclosed, and the preparation method is the patent’s Example 1) The culture medium replaces the medium for 3 days, and the one without the compound is set as a negative control, and the exocrine is not added. The body without compound was regarded as the blank control, and only the culture m...

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Abstract

The invention relates to application of a combination of polypeptide conjugates and epidermal stem cell exosomes in medicine and cosmetics. According to the present invention, exosomes are extracted and obtained from epidermal stem cells, the exosomes and polypeptide conjugates are prepared into a composition that inhibits melanin synthesis, and at the same time, skin experiments prove that the composition has the effects of removing freckles and preventing acne.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of a combination of polypeptide conjugates and epidermal stem cell exosomes in medicines and cosmetics. Background technique [0002] Exosomes are subcellular double-layer membrane vesicles with molecular diameters of 40-100 nm that are formed and secreted by cells through a series of regulatory processes such as "endocytosis-fusion-efflux". Under the transmission electron microscope, exosomes are flat or spherical, and their density in sucrose solution is 1.13-1.19 g / mL. The density is related to the cell source and changes with the protein content. Exosomes do not contain DNA fragments, but contain proteins such as cytokines and growth factors similar to their source cells, as well as biologically active substances such as lipids, coding or non-coding RNA, which play an important role in regulating the physiological functions of cells. The plasma membrane...

Claims

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Application Information

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IPC IPC(8): A61K35/36A61K47/64A61P17/00A61P17/02A61K8/99A61K8/98A61K8/64A61K8/49A61Q19/02A61K31/5377
CPCA61K8/49A61K8/64A61K8/985A61K8/99A61K31/5377A61K35/36A61K47/641A61P17/00A61P17/02A61Q19/02A61K2300/00
Inventor 彭菲李静
Owner 广州市新纪元化妆品有限公司
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