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Genetically modified mesenchymal stem cell extract, extraction method and application in skin tightening and aging resistance

A stem cell and genetic modification technology, applied in the field of genetic engineering, can solve problems such as trauma and affect the appearance of the skin, and achieve the effect of improving skin tone and texture

Inactive Publication Date: 2021-01-01
SHANDONG XINRUI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this treatment is invasive and carries certain risks, causing a lot of trauma and may affect the appearance of the skin
In addition, the application of this traditional cell therapy has many limitations, because live cells are used in this therapy, and the safety of live cells should be strictly evaluated before injection

Method used

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  • Genetically modified mesenchymal stem cell extract, extraction method and application in skin tightening and aging resistance
  • Genetically modified mesenchymal stem cell extract, extraction method and application in skin tightening and aging resistance

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The secreted Klotho gene of modified mesenchymal stem cells, the nucleic acid sequence of which is shown in SEQ ID NO.1. The pAV-EF1α-Klotho plasmid was obtained by plasmid preparation technology, and after being sequenced correctly by Shanghai Sangon Bioengineering Co., Ltd., a large amount was extracted for use.

Embodiment 2

[0043] Preparation of umbilical cord mesenchymal stem cells

[0044] Take the newborn umbilical cord donated by the hospital, disinfect it twice with 75% alcohol in the ultra-clean workbench, put it in a petri dish, use tweezers to peel off the Fahrenheit jelly tissue, and cut it to 0.5mm with scissors 2 sized nubs. Transfer the shredded Wahrenheit jelly tissue to a culture bottle, add UltraCULTURE medium containing plasma substitute for culture, and observe with a microscope every day. When the stem cells that climbed out covered 80% of the bottom of the bottle, they were subcultured. After subculture, the growth rate of the cells was accelerated, and they were passed to the P4 generation for experiments every 2-3 days.

Embodiment 3

[0046] The 293T cells were revived by conventional techniques, and used for transfection experiments when the culture was in good condition. Mix pAV-EF1α-Klotho, packaging plasmid, and helper plasmid at a ratio of 1:1:1, transfect 293T cells in good condition, 37°C, 5% CO 2 Cultivate in an incubator for 48 hours, collect the supernatant, centrifuge at 2000 g for 10 minutes at 4°C, transfer to a new EP tube, filter with a 4.5 μm filter, and store at -80°C. Carry out virus titer determination, the virus titer that obtains among the present invention is 3.3 * 10 7 TU / mL.

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Abstract

The invention discloses a genetically modified mesenchymal stem cell extract, an extraction method and application in skin tightening and aging resistance. The genetically modified mesenchymal stem cell extract at least contains one of SDF-1, VEGF, CXCR4 and klotho, and a detection method comprising the following steps is adopted for verification: extracting total RNA of cells; removing genomic DNA; performing reverse transcription reaction; digesting and lysing genetically modified mesenchymal stem cells to obtain the genetically modified mesenchymal stem cell extract. The genetically modified mesenchymal stem cell extract can promote expression of hyaluronic acid, elastin, collagen or reticulamin in the skin.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a genetically modified mesenchymal stem cell extract, an extraction method and its application in skin tightening and anti-aging. Background technique [0002] In human life, improving physical appearance occupies a larger focus than all other daily life-related issues combined. There are far more consumer products that beautify the human body than those that treat human diseases. Improving the tone and appearance of the body is a growing multi-billion dollar industry that includes cosmetics, nutritionals, pharmaceuticals and physical therapy. Consumer attention is focused on the latest ingredients for anti-aging, anti-wrinkle, body contouring, varicose vein reduction, acne elimination and weight loss combinations. Among all these cosmetic beauty concerns, the improvement of facial appearance is the biggest concern and concern of human beings, especially women. The...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/99A61K8/98A61Q19/08A61Q19/02A61Q19/00A61K35/28A61P17/18A61P17/00C12N5/10C12N15/861
CPCA61K8/99A61K8/98A61Q19/08A61Q19/02A61Q19/00A61K35/28A61P17/18A61P17/00C12N5/0668C12N15/86C12N9/2402C12Y302/01031C12N2510/00C12N2710/10043C12N2800/107
Inventor 刘明录王立新韩庆梅万磊冯建海金海锋强邦明卢永灿刘媛媛
Owner SHANDONG XINRUI BIOTECH CO LTD
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