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Application of transgenic stem cell secretion in preparation of cosmetics

A stem cell and stromal cell technology, which is applied to the secretion of transgenic stem cells in the field of cosmetic preparation, can solve problems such as excessive melanin deposition, and achieve the effects of inhibiting proliferation, inhibiting melanin production, and inhibiting protein expression.

Inactive Publication Date: 2022-02-01
苗晓明
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on the use of exosomes in whitening drugs or cosmetics. Therefore, studying the use of exosomes in whitening will help to further solve the problem of excessive melanin deposition

Method used

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  • Application of transgenic stem cell secretion in preparation of cosmetics
  • Application of transgenic stem cell secretion in preparation of cosmetics
  • Application of transgenic stem cell secretion in preparation of cosmetics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Construction of overexpression vector of non-coding gene LNC-RP11-452J13.1

[0035] (1) Amplification primers were designed according to the transcript sequence ENST00000420807.1 (SEQID NO.1) of the non-coding gene LNC-RP11-452J13.1. The plasmid used was pcDNA3.1, and the enzymes were XbaI and EcoRI enzymes. The designed primers The sequence is as follows:

[0036] Upstream primer: 5'-TCTAGACAATGATTCCATTATCA-3', SEQ ID NO.2;

[0037] Downstream primer: 5'-GAATTCTTGATTGAGAGCCCGTG-3', SEQ ID NO.3;

[0038] (2) Bone marrow mesenchymal stem cells were inoculated on cell culture plates, the control group was transfected with pcDNA3.1, and the experimental group was transfected with pcDNA3.1-LNC-RP11-452J13.1;

[0039] (3) After 48 hours of transfection, add Trizol and extract RNA according to the instructions;

[0040] (4) Reverse transcribe RNA into cDNA according to the reverse transcription kit;

[0041] (5) Detect the relative expression of the non-coding gene LNC-RP...

Embodiment 2

[0046] Obtaining exosomes from bone marrow mesenchymal stem cells transfected with pcDNA3.1-LNC-RP11-452J13.1

[0047] (1) Bone marrow mesenchymal stem cells transfected with pcDNA3.1 and pcDNA3.1-LNC-RP11-452J13.1 were inoculated into culture flasks;

[0048] (2) When the cells enter logarithmic growth, collect the medium in the culture flask, centrifuge at 2000r / min at 4°C for 10min, collect the supernatant and filter it with a 0.22μm filter;

[0049] (3) After filtering, add the culture medium back into the culture bottle and add 1ml of fresh culture medium;

[0050] (4) Change the medium once a day according to the above method. After culturing for 72 hours, collect the medium in a centrifuge tube and filter it with a 0.22 μm filter;

[0051] (5) Centrifuge the medium at 300×g at 4°C for 10 minutes to remove residual cells, collect the supernatant, and remove the precipitate;

[0052] (6) Centrifuge the medium at 2000×g for 10 minutes at 4°C to remove dead cells, collect...

Embodiment 3

[0057] Detect the exosome extraction effect and the expression level of LNC-RP11-452J13.1 in the exosomes of the experimental group and the control group

[0058] 1. Detection of protein expression of exosome markers in cell extraction and exosome extraction proteins

[0059] (1) Extract the cell lysed proteins of exosome 1 and exosome 2, and prepare protein samples;

[0060] (2) Prepare 10% separating gel and 5% stacking gel electrophoresis gel, install the prepared electrophoresis gel into the electrophoresis tank, and use a pipette to add samples;

[0061] (3) Use 80V power supply to run the separation gel and stacking gel throughout the process, and stop electrophoresis when the bromophenol blue reaches the bottom of the glass plate;

[0062] (4) Pre-cut the PVDF membrane for use, soak the PVDF membrane in methanol for 5-10 minutes before transferring the membrane, and make a sandwich model of electroporation, electroporation at 220mA for 1.5h;

[0063] (5) After the mem...

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Abstract

The invention provides application of transgenic stem cell secretion in preparation of cosmetics, and belongs to the technical field of biology. The transgenic stem cell secretion is an exosome of a bone marrow mesenchymal stem cell overexpressing LNC-RP11-452J13.1 gene, wherein the exosome can significantly inhibit the proliferation of melanoma cells B16, can significantly inhibit the protein expression of tyrosinase, and can significantly inhibit the generation of melanin.

Description

technical field [0001] The invention belongs to the technical field of biology, and in particular relates to the use of transgenic stem cell secretion in the preparation of cosmetics. Background technique [0002] With the rapid development of the contemporary economy, environmental pollution is becoming more and more serious. Environmental pollution causes the hole in the ozone layer to expand, resulting in more and more ultraviolet rays that the human skin contacts and absorbs, which will cause melanocytes to synthesize melanin. Disability. Melanin is a pigment widely distributed on the surface of the skin, retina and hair. Melanin is synthesized from tyrosine through the enzymatic oxidation of Tyr. The production of melanin is one of the body's main defense mechanisms against the sun's UV rays. Ultraviolet rays, chronic inflammation, and hormone stimulation can cause the skin to produce melanin, but when melanin is abnormally produced, problems such as skin pigmentatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/99A61K8/98A61Q19/02A61K35/28A61P17/00C12N5/10C12N15/12C12N15/85
CPCA61K8/99A61K8/981A61Q19/02A61K35/28A61P17/00C12N5/0663C12N15/85C07K14/47A61K2800/782C12N2510/00
Inventor 苗晓明郭城
Owner 苗晓明
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